Multiple Oblique Illumination and High-Definition Microscopy for Breast Fine-Needle Aspirates

ABSTRACT We wished to determine whether small-intestinal submucosa (SIS) will epithelialize when used as a ureteral replacement material. An 11-mm segment of native ureter was excised from eight New Zealand White rabbits and replaced with an 11-mm porcine SIS graft, which was circumferentially wrapped around a ureteral stent. The SIS ureteral grafts were harvested at 11 days or 35 days postimplantation and examined grossly and by standard light microscopy techniques. Partial epithelialization with the ingrowth of urothelium, smooth muscle cells, and blood vessels was observed in the grafts harvested at 11 days postimplantation. The SIS ureteral grafts examined at 35 days postimplantation showed additional restructuring of the smooth muscle cell layer and more organized epithelialization in comparison to the SIS graft examined at 11 days. After 35 days of regenerative healing, elements of all three layers of the native ureter were observed within the collagen matrix of the SIS graft. No significant complications were observed, but all subjects (8/8) demonstrated mild intra-abdominal adhesions. Mild collecting system dilatations were observed in 4/4 (100%) of the animals harvested at 35 days and in 0/4 (0%) of the animals harvested at 11 days. We have this demonstrated in this preliminary study that SIS xenografts will epithelialize when used as a ureteral replacement material. The repair mechanism of these ureteral grafts occurred through a regenerative healing process rather than by scar formation. With further studies, this material may prove to be a useful treatment option in patients with ureteral injuries.     

Ureteral segment replacement using a circumferential small-intestinal submucosa xenogenic graft.

We wished to determine whether small-intestinal submucosa (SIS) will epithelialize when used as a ureteral replacement material. An 11-mm segment of native ureter was excised from eight New Zealand White rabbits and replaced with an 11-mm porcine SIS graft, which was circumferentially wrapped around a ureteral stent. The SIS ureteral grafts were harvested at 11 days or 35 days postimplantation and examined grossly and by standard light microscopy techniques. Partial epithelialization with the ingrowth of urothelium, smooth muscle cells, and blood vessels was observed in the grafts harvested at 11 days postimplantation. The SIS ureteral grafts examined at 35 days postimplantation showed additional restructuring of the smooth muscle cell layer and more organized epithelialization in comparison to the SIS graft examined at 11 days. After 35 days of regenerative healing, elements of all three layers of the native ureter were observed within the collagen matrix of the SIS graft. No significant complications were observed, but all subjects (8/8) demonstrated mild intra-abdominal adhesions. Mild collecting system dilatations were observed in 4/4 (100%) of the animals harvested at 35 days and in 0/4 (0%) of the animals harvested at 11 days. We have this demonstrated in this preliminary study that SIS xenografts will epithelialize when used as a ureteral replacement material. The repair mechanism of these ureteral grafts occurred through a regenerative healing process rather than by scar formation. With further studies, this material may prove to be a useful treatment option in patients with ureteral injuries.     

Use of a composite skin graft composed of cultured human keratinocytes and fibroblasts and a collagen-GAG matrix to cover full-thickness wounds on athymic mice

In patients with extensive full-thickness burns, wound coverage may be accelerated if skin can be expanded to produce a skin replacement that reproducibly supplies blood to the wound and has good structural qualities. In addition, development of skin replacements may benefit patients who require reconstruction or replacement of large areas of abnormal skin. We have developed a composite skin replacement composed of cultured human keratinocytes (HK) and fibroblasts. Cultured human fibroblasts are seeded into the interstices, and cultured HKs are applied to the surface of a matrix composed of type I collagen crosslinked with a glycosaminoglycan, which has a defined physical structure. After HKs reach confluence on the matrix surface, the composite grafts are placed on full-thickness wounds on the dorsum of athymic mice. Graft acceptance, confirmed by positive staining with antibodies specific for human HLA-ABC antigens on HKs, is approximately 90%. A defined skin structure is present histologically by day 10 after grafting, with a differentiated epithelium and a subepidermal layer densely populated by fibroblasts and capillaries without evidence of inflammation. Fluorescent light microscopy to identify laminin and type IV collagen and electron microscopy confirm the presence of basement membrane components by 10 days after grafting. Attachment of the graft to the wound is similar with and without the addition of human basic fibroblast growth factor, a potent angiogenic agent, to the skin replacement before graft placement on wounds.     

First prize: ureteral segmental replacement revisited

BACKGROUND AND PURPOSE: Long strictures of the proximal ureter are difficult to manage, and circumferential replacement with various natural and synthetic materials has been unsuccessful. We sought to use cultured autologous cells seeded onto graft material for proximal-ureteral replacement. Additionally, we wished to determine if urothelial cell-seeded de-epithelialized small bowel would generate adequate ureteral replacement. MATERIALS AND METHODS: Three sets of experiments were performed. First, autologous pig-bladder smooth-muscle and urothelial cells were expanded in culture on large sheets of multilayer small-intestinal submucosa (SIS). These sheets were then tubularized and used to replace a 5-cm segment of proximal ureter in pigs. Second, autologous cells harvested from the bladders of Beagle dogs were cultured and seeded on porcine ureteral acellular matrix, which was used to replace a 3-cm segment of ureter in dogs. Segments were wrapped in omentum to enhance vascularity. Third, a de-epithelialized small-bowel segment seeded with autologous bladder-epithelial cells was transversally retubularized (Monti) into a 4-cm ureteral replacement. Follow-up studies consisted of retrograde pyelography, serum chemistry assays, hematoxylin/eosin studies, and immunohistopathologic examination using antibodies against alpha-smooth-muscle actin and pancytokeratin AE1-AE3. RESULTS: Coculture of urinary-tract cells on large segments of SIS failed to create adequate ureteral replacement. All grafts were contracted and stenotic, with complete obstruction of the ipsilateral renal unit. Similar results were seen in the Beagles. Despite clinical obstruction and gross contraction of the graft, a circumferential muscular ureteral wall lined with multilayer transitional epithelium was present. Urotheliumseeded de-epithelialized Monti bowel segments resulted in patent ureteral replacement without hydroureteronephrosis and with normal renal function, serum electrolytes, and acid-base balance. However, bowel mucosa fully regenerated, with multilayer transitional epithelium growing adluminally in continuity with the proximal and distal anastomotic sites. CONCLUSIONS: Seeding of ureteral grafts with autologous bladder cells does not promote success in two largeanimal models using different xenogenic acellular matrices. However, muscle and urothelium regeneration occurs with ureteral acellular matrix in the dog. Urothelium-seeded de-epithelialized Monti bowel segments may be an acceptable substitute for long proximal ureteral segments. Further technical refinements are required to replace the bowel mucosa completely with normal urothelium.     

Canine ureteral replacement with long acellular matrix tube: is it clinically applicable?

PURPOSE: We evaluated the effectiveness of acellular matrix used as a tube for replacement of a relatively long segment of the canine ureter. MATERIALS AND METHODS: Acellular matrix was obtained by excision of the whole ureter of donor dogs that were sacrificed and not included in the study group. Retrieved ureters were treated to have complete cell lysis, while maintaining the fiber framework. The study included 10 mongrel dogs in which a 3 cm segment was excised from 1 ureter and replaced by a tube of acellular matrix of the same length and width. The new tube was sutured proximal and distal by watertight interrupted sutures around a 5Fr Double-J stent (Medical Engineering Corp., New York, New York) that remained for 6 weeks. Excretory urography was done 1 and 2 weeks after stent removal and the dogs were then sacrificed. Before sacrifice the ureter was exposed and carefully examined, and the whole specimen was excised for histopathological examination. RESULTS: All dogs survived surgery except 1, which died 1 week postoperatively of a malpositioned stent and urinary ascites. There was no clinically apparent postoperative complications during the presence or after the removal of the ureteral stents. One week after stent removal excretory urography showed ipsilateral mild to moderate hydroureteronephrosis in 3 dogs and no dye excretion in 6 with a normal contralateral kidney. One week later no dye excretion was detected in all except 1 dog, which showed more radiological deterioration. At the time of sacrifice there was moderate to marked hydroureteronephrosis above the level of the new tube in all dogs. Although the graft was intact in all subjects, marked shrinkage was observed. On ureteral calibration there was significant narrowing of the lumen up to complete occlusion. At 8 weeks histopathological examination showed extensive fibrosis. CONCLUSIONS: An acellular matrix tube is not able to replace a 3 cm segment of the canine ureter.     

Small Intestinal Submucosa as a Small-Diameter Arterial Graft in the Dog

Autogenous saphenous vein, human umbilical vein, modified bovine collagen, Dacron, and PTFE have been used as small-diameter arterial grafts with moderate success. We tested autogenous small intestine submucosa as a small-diameter arterial graft in both a carotid and femoral artery (mean ID 4.3 mm) of 18 dogs (total of 36 grafts). All dogs received aspirin and warfarin sodium for the first 8 weeks after surgery. Graft patency was evaluated by Doppler ultrasound techniques and angiography. Two grafts ruptured and 5 grafts occluded by 21 days after surgery. One graft became occluded at 14 weeks. Fifteen dogs were sacrificed at periodic intervals until 48 weeks after surgery. Patent grafts had no evidence of infection, propagating thrombus, or intimal hyperplasia. Graft aneurysmal dilation occurred in 4 grafts (11%). The grafts were composed of a dense organized collagenous connective tissue with no evidence of endothelial cell growth on the smooth luminal surface. Three dogs are alive at 76 to 82 weeks after surgery. Overall, graft patency was 75%. Graft patency after cessation of anticoagulation therapy was 92.3% (12 of 13 grafts). We conclude that autogenous small intestinal submucosa can be used as a small-diameter arterial graft in the dog and is worthy of further investigation.     

Small intestinal submucosa as a small-diameter arterial graft in the dog

Autogenous saphenous vein, human umbilical vein, modified bovine collagen, Dacron, and PTFE have been used as small-diameter arterial grafts with moderate success. We tested autogenous small intestine submucosa as a small-diameter arterial graft in both a carotid and femoral artery (mean ID 4.3 mm) of 18 dogs (total of 36 grafts). All dogs received aspirin and warfarin sodium for the first 8 weeks after surgery. Graft patency was evaluated by Doppler ultrasound techniques and angiography. Two grafts ruptured and 5 grafts occluded by 21 days after surgery. One graft became occluded at 14 weeks. Fifteen dogs were sacrificed at periodic intervals until 48 weeks after surgery. Patent grafts had no evidence of infection, propagating thrombus, or intimal hyperplasia. Graft aneurysmal dilation occurred in 4 grafts (11%). The grafts were composed of a dense organized collagenous connective tissue with no evidence of endothelial cell growth on the smooth luminal surface. Three dogs are alive at 76 to 82 weeks after surgery. Overall, graft patency was 75%. Graft patency after cessation of anticoagulation therapy was 92.3% (12 of 13 grafts). We conclude that autogenous small intestinal submucosa can be used as a small-diameter arterial graft in the dog and is worthy of further investigation.     

Experimental aortic replacement with a vascularized tissue graft

Prosthetic graft infection in the aortic position remains a catastrophic complication of aortic replacement. In an effort to incorporate the advantages of vascularized tissue into aortic replacement, the practionally of infrarenal aortic replacement was investigated in dogs by a vascularized, musculofascial, pedicle graft. Eight dogs underwent laparotomy, and the left rectus abdominus muscle, its posterior rectus sheath, and underlying peritoneum were used to construct a tube graft based on the inferior epigastric vascular pedicle. A 2- to 4-cm section of the infrarenal aorta was resected, and the tube graft was interposed. Aortography documented graft patency immediately and at 1,6, and 10 months. Gross and histologic examination of the grafts at one month revealed viable skeletal muscle in the pedicle graft, with a thin layer of fibrin deposition on the luminal surface. Ten months after insertion, the microscopic examination against revealed patent vessels in the graft and a neoendothelial layer that lined the luminal surface of the graft. Successful replacement of the dog infrarenal aorta can be performed using a tube graft as part of a vascularized, musculofascial, pedicle flap. The potential for its use in the infected field is intriguing and deserves further study.     

Infection of vascular prostheses caused by bacterial biofilms

A canine model was developed to study the efficacy of graft replacement as treatment for vascular prosthesis infections from Staphylococcus epidermidis. Infrarenal aortic graft infections were established in 18 dogs by implantation of Dacron prostheses colonized in vitro with a slime-producing strain of S. epidermidis to form an adherent bacteria-laden biofilm (5 X 10(6) colony-forming units/cm2 graft). Study animals developed a graft infection with anatomic and microbiologic characteristics typical of late prosthetic graft infections in humans (sterile perigraft exudate, absent graft incorporation, and normal serum leukocyte count and sedimentation rate). The S. epidermidis study strain was isolated from 14 of 18 explanted grafts (78%) by mechanical disruption of the graft surface biofilm and culture in broth media. Four dogs with sterile graft cultures had histologic evidence of bacterial infection. The established prosthetic surface biofilm infection was treated by graft excision, parenteral cefazolin, and graft replacement with a Dacron or polytetrafluoroethylene (PTFE) vascular prosthesis. One month after graft replacement, no PTFE graft had signs of infection, but perigraft exudate and inflammation involved three of nine Dacron grafts (33%). The study strain was recovered from four of nine PTFE grafts (44%) and two of nine Dacron (22%) replacement grafts (p greater than 0.05). Prosthetic replacement of Dacron prostheses infected by S. epidermidis as a bacteria-laden surface biofilm can result in early graft healing, but persistent colonization of one third of replacement grafts signify that recurrent clinical infection remains a risk.     

涤纶血管胶原蛋白涂层的实验及临床研究

提取牛皮Ⅰ型胶原蛋白,以4mg/ml的胶原蛋白溶液涂层苏州涤纶人工血管,经过0.3％戊二醛交联,涤纶血管渗水量由383±28ml·min~(-1)/cm~2降至2.8±0.5ml·min~(-1)/cm~2。将胶原蛋白涂层涤纶血管移植犬腹主动脉下段,以白蛋白涂层涤纶血管移植为对照。在肝素化状态下均不渗血。术后30天和90天观察显示,胶原涂层涤纶血管内壁覆盖一层完整光滑的内膜,光镜显示为纤维结缔组织,表面有内皮细胞被覆并与自体动脉内膜延续。而白蛋白涂层涤纶血管血栓形成率高,内壁纤维素沉积。临床采用胶原涂层涤纶血管为4例马凡综合征病人行主动脉根部替换,术中涤纶血管不渗血,随访3个月无不良反应。本研究显示胶原涂层涤纶血管机械性能良好,不渗血,尤其能促进涤纶血管的愈合。作为主动脉移植材料可广泛应用于临床,若用于中小血管移植可望提高远期通畅率。     

Development of a small caliber vascular graft using a connective tissue tube with temporary antithrombogenicity

A connective tissue tube containing collagen fibrils and capillary blood vessels is an ideal material for artificial artery, since the collagen fibrils compose an excellent supporting framework for cell migration and proliferation and the endothelial cells of the capillary can be the origin of the endothelialization of the graft. However, its thrombogenicity has remained a problem. This experiment was designed to develop a small caliber (3mm) vascular graft using an autogenous connective tissue provided with temporary antithrombogenicity. A tube knitted of ultra-fine polyesters was implanted subcutaneously in the back of mongrel dogs for three weeks. Thus accomplished connective tissue grafts were heparinized ionically in vitro, using cross-link agents. Then the treated grafts, 5-6 cm in length and 3mm in diameter, were segmentally replaced with bilateral carotid arteries. During the follow-up period up to 55 days, 7 of 8 grafts (88%) were patent. Histological examinations revealed that there was no thrombus and the neointima formation with a lining of endotherial cells was completed, extending overall length of the grafts. Thus, newly devised hybrid type of connective tissue graft is thought to be usefull for a replacement for arteries with a small caliber.     

Persistent platelet activation by passivated grafts.

Vascular grafts in canines exhibit similar healing patterns to humans in that the graft surface forms a pseudointima over time but endothelializes only near the anastomotic sites. Thus the pseudointima at the midportion of the graft may represent a nidus for persistent platelet activation. The purpose of this investigation was to examine the effect of the maturing graft surface on platelet activation. Long Dacron subcutaneous carotid to aorta grafts (50 cm x 8 mm) were placed in nine dogs. Blood samples were obtained by direct graft puncture, at the proximal and distal ends of the graft, at 1, 24, 48, 72 hours, 1, 2, 3, 4 weeks, and monthly thereafter for 8 months. Seven sham dogs had subcutaneous grafts implanted without arterial anastomoses, and blood samples were drawn from the femoral artery. Platelet counts were determined with a platelet counter. Platelet aggregation and release of adenosine triphosphate was determined with a whole blood aggregometer by use of arachidonic acid, collagen, and adenosine diphosphate as agonists. No difference was found in platelet aggregation to collagen or adenosine diphosphate stimulation across the graft, but platelets released significantly less adenosine triphosphate to collagen and adenosine diphosphate stimulation distally versus proximally. In the graft dogs a decrease in systemic platelet counts of 50% occurred from the preoperative level which persisted over 8 months (p less than 0.01). Also less response occurred to collagen and ADP stimulated platelet aggregation in the graft animals than the sham animals during the first month of study. These data suggest that significant platelet-graft interactions occur even after the graft has formed a mature pseudointima.     

Reconstruction of ureteral defects with microvascular vein grafts in a rat model

This study was undertaken to test the performance of an autologous vein graft as a ureteral replacement in the rat model. Twenty-six rats were divided into three groups. In Group 1 (n = 10), the animals had a 3-mm segment of the ipsilateral ureter excised and the ureteral defect repaired, using a superficial epigastric vein graft. In Group 2 (n = 10), the same ureteral defect was created and again repaired, using a superficial epigastric vein graft, with the addition of a Silastic stent. The control, Group 3 (n = 6), had the ureter transected and repaired solely by means of primary anastomosis. Animals from each group underwent urography and were sacrificed for histology at three different postoperative intervals: 1, 4, and 12 weeks. Postoperative urography results showed normal renal function in the animals with ureteral reconstruction using vein grafting aided by a stent, as well as in those with primary ureteral anastomosis. No renal function return was seen in the animals with ureteral reconstruction by vein grafting alone, without stent support. Histologically, a progressive loss of the vascular endothelium, and replacement with the urothelium typical of the ureter, was seen in the stented vein grafts. Severe ureteral obstruction at the proximal site of the graft and hydronephrosis were seen in the vein-graft group without stenting. This study demonstrates that autologous vein grafts can be used successfully to correct a ureteral deficit, contingent on accurate microsurgical technique and immediate stenting.     

Use of an antibiotic-bonded graft for in situ reconstruction after prosthetic graft infections.

We have developed an infection resistant vascular prosthesis by bonding rifampin to Dacron grafts with the use of a collagen matrix release system. The purpose of this study was to determine the efficacy of this antibiotic-bonded graft in resisting infection after an in situ reconstruction of a previously infected prosthetic bypass. Eighty-three adult mongrel dogs underwent implantation of a 3 cm untreated Dacron graft into the infrarenal aorta. This initial graft was deliberately infected, at the time of operation, with 10(2) organisms of Staphylococcus aureus by direct inoculation. One week later, the dogs were reexplored, the retroperitoneum debrided, and the animals randomized to undergo an end-to-end in situ graft replacement with either one of two types of prosthetic grafts: group I (collagen, n = 36) received control collagen-impregnated knitted Dacron grafts; group II (rifampin, n = 47) received experimental collagen-rifampin-bonded Dacron grafts. Each group of animals was then subdivided to receive one of four treatment protocols: (a) no antibiotic therapy, (b) cephalosporin peritoneal irrigation solution (cefazolin 500 mg/1000 ml) during operation and two doses of cephalosporin (cefazolin, 500 mg intramuscularly) postoperatively, (c) treatment as in protocol group b plus 1 week of cephalosporin (cefazolin, 500 mg intramuscularly, twice daily), and (d) treatment as in protocol group b plus 2 weeks of cephalosporin (cefazolin, 500 mg intramuscularly, twice daily). All grafts were sterilely removed between 3 and 4 weeks after implantation. There were no anastomotic disruptions and all grafts were patent at the time of removal.(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphologic changes in the canine urinary tract after ureteral reconstruction with the tunica albuginea (experimental study)*

Objectives: The problem of ureteral reconstruction has not been fully solved, despite dramatic developments in surgical methods. Many attempts to repair a ureteral damage with autologous or homologous organ grafts, or with organic substitutes have not really been satisfactory. Also, attempts to use alloplastic and heteroplastic prostheses made of synthetic materials have been equally disappointing. Experimental studies, particularly in dogs, indicate great regenerative abilities of ureters when appropriate surgical techniques are used. Materials and methods: In our experimental study, suitability of the tunica albuginea for reconstruction of the middle part of the ureter was investigated in 20 dogs. The dogs were divided into two groups according to a type of ureteral damage: partial (a patch graft) or complete (a tube graft). The ureteral damage was 2–3 cm long in 10 dogs and 3–3.5 cm long in the subsequent 10 dogs. Each dog underwent two operations. The first operation involved a fenestrating incision in the ureter, 23.5 cm long, so as to preserve the ureteral continuity, and then the produced injury was repaired with a patch of the tunica albuginea, with the 4 Charr catheter being inserted in the ureter. The second operation involved a complete excision of ureteral segments, 2–3.5 cm long, on the contralateral side. The excised length of the ureter was substituted with a tube graft of the tunica albuginea, also using the 4 Charr catheter. Thus, in 21 cases, a partial damage of the ureteral wall was repaired with a patch graft of the tunica albuginea. In 19 cases, a complete damage of the ureter was repaired with a tube graft of the tunica albuginea. Results: Four dogs died after the tube-graft implantation due to urinary infiltration and/or purulent inflammatory changes in the kidneys. Autopsies were performed in all the dogs after they had been put to sleep. Definitely poor results were obtained in case of the complete reconstruction of the ureteral loss with a tube graft of the tunica albuginea, 3–3.5 cm long. Within the graft area, we observed inflammatory infiltrates of varied intensity, fibrosis, cicatricial strictures or ureteral obstruction. Occasionally, necrotic foci, degenerative changes or hydronephrosis were found. In case of the reconstruction with a shorter tube graft, i.e. 2–3 cm long, similar changes were observed, however, they were less intense. Satisfactory results were obtained following the partial reconstruction of the ureteral loss with a patch graft of the tunica albuginea. In this case, we observed generally normal regeneration not only of the transitional epithelium but the muscular layer and ureteral structure as well, with full patency of the ureter preserved. The findings obtained in our study are compatible with the findings of other authors. Conclusions: (1) The success of reconstructive procedures in the ureters can be achieved if the continuity of the ureter is preserved and the catheter is removed after 14 days. (2) The substitution of a complete ureteral loss with a tube-graft of the tunica albuginea results in necrosis of the graft and restenosis. *Printed in part at the postdoctoral thesis of Tyloch Ferdynand MD: “An experimental study on the use of tunica albuginea for supplying at westage of ureters” (Badania doswiadczalne nad przydatnoscia blony bialawej jadra w operacjach plastycznych moczowodu) University School of Medical Sciences Bydgoszcz 1994.     

Small intestinal submucosa as a large diameter vascular graft in the dog

Autogenous saphenous vein and synthetic materials, such as Dacron and expanded polytetrafluoroethylene, have been used extensively as vascular grafts with moderate success. Improved success rates for vascular graft surgery may be possible if superior graft material was available. We tested the use of autogenous small intestinal submucosa (SIS) as a large diameter (10 mm) vascular graft in the infrarenal aorta of 12 dogs. One dog died with graft thrombosis within 48 hr of surgery. Nine dogs were sacrificed at various times during a 52-week post-surgical period and showed patent grafts without infection, thrombosis, intimal hyperplasia, or adverse effects upon blood pressure. There was no ultrastructural evidence of endothelial cell growth on the luminal surface of the SIS graft which was composed of a dense, non-thrombogenic, organized collagenous connective tissue. The SIS material was approximately one order of magnitude less elastic than natural aorta and showed an immediate dilatation of approximately 18% after exposure to the systemic blood pressure. However, there was no progressive dilatation during the 52-week postsurgical period. Two dogs remain alive at 8 and 52 weeks post-surgery with patent grafts as determined by positive contrast radiography and Doppler studies. We conclude that autogenous small intestinal submucosa can be successfully used as a large diameter arterial graft in the dog and is worthy of further investigation.     

Gore-Tex Grafts for Replacement of the Superior Vena Cava

In an effort to evaluate polytetrafluoro-ethylene (Gore-Tex) as a replacement for large veins, tubular grafts of the material were substituted for the precava in 33 mongrel dogs weighing 15 to 30 kg. Thirteen dogs had grafts 10 cm long by 12 mm wide with a 15- to 30-u pore size; 15 dogs had grafts of the same length and width from 90-μ. pore size material; and 5 dogs had grafts 5 cm long by 12 mm wide, of 90-μ pore size Gore-Tex. The effect of Gore-Tex tubes with and without spiral support and the differences in anatomical positions of the grafts were evaluated in terms of patency. Dogs were killed if a venogram showed occlusion of the graft. Twelve dogs survived 90 days; the longest survivor was killed at eleven months. At postmortem examination, the graft was patent but extremely narrow at the atrial end. Modification in the fabrication of Gore-Tex may eliminate factors contributing to graft failure; experiments longer than 90 days are necessary to evaluate Gore-Tex as a large vein replacement material. The 90-μ pore size material used in this experiment was unsuitable as a canine venous substitute.     

A histologic assessment of canine anterior cruciate substitution with bovine xenograft

Glutaraldehyde cross-linked bovine xenograft anterior cruciate ligament replacement has been investigated in dogs. The implants were functional for the activities of the dogs at one year of follow-up study. The knees were stable, and the articular surfaces on gross inspection showed only minimal evidence of degenerative changes. Histologic evaluation of the implants demonstrated a progressive ingrowth of the xenograft by host tissues. The host tissues grew parallel to the collagen scaffolding of the graft and consisted of host blood vessels and fibroblastic elements that produced collagen of host origin. The host response was benign in appearance without cellular evidence of rejection phenomena.     

Wound healing effect of acellular artificial dermis containing extracellular matrix secreted by human skin fibroblasts

In this study, an acellular artificial dermis, composed of human collagen and glycosaminoglycan (GAG) secreted by cultured human fibroblasts on a bovine collagen sponge, was developed. Much of the newly secreted extracellular matrix (ECM) remained after the cell removal process. The main theme of this study focused on the matrix, rather than the viable cell components of the skin, as the major dermal deficit in the wound. Both the acellular artificial and bioartificial dermises, containing viable cells with ECM, were significantly less soluble than the collagen sponge, and the relative GAG content in the bioartificial and acellular artificial dermises was approximately 115-120% of the chondroitin-6-sulfate (CS) content found in the collagen sponge. In the group receiving the collagen sponge, the wound area gradually decreased to approximately 10% of its original area, while in the groups receiving the bioartificial and acellular artificial dermises, the wound area also gradually decreased to approximately 60 and 50%, respectively, of the original size over the 5 weeks after grafting. Both the bioartificial and acellular artificial dermises formed thicker, denser collagen fibers; more new blood vessel formation was observed in both cases. The basement membrane of the regenerated epidermal-dermal junction was thicker and more linear in the acellular artificial dermis graft than in the collagen sponge graft. In conclusion, the wound healing effects of acellular artificial dermis are no less than those of the bioartificial dermis, and much better than the collagen sponge graft with respect to wound contraction, angiogenesis, collagen formation, and basement membrane repair.     

Evaluation of a xenogeneic acellular collagen matrix as a small-diameter vascular graft in dogs--preliminary observations.

Autogenous veins are the materials of choice for arterial reconstruction. In the absence of autogenous material, prosthetic materials are used. However, vascular prostheses of less than 0.4 cm in diameter have low long-term patency. This study was designed to determine if cells would infiltrate an engineered xenogeneic biomaterial used as a small diameter arterial graft in dogs and, if so, to determine the phenotype of the infiltrating cells. Nine acellular xenogeneic grafts (0.4 cm in diameter, 5 cm long), composed of porcine collagen derived from the submucosa of the small intestine and type I bovine collagen, were implanted as end to-end interposition grafts in femoral arteries of five male mongrel dogs (total of nine grafts). All dogs received daily aspirin (325 mg). Patency of implanted grafts was monitored weekly by Duplex ultrasonography. After 9 weeks, or earlier in case of blood flow reduction by at least 75%, grafts were explanted and prepared for light or electron microscopy to evaluate cellularization. Eight of nine grafts remained patent up to 9 weeks. At explant, diameters were 0.31 +/- 0.02 cm at the midgraft, and 0.14 +/- 0.01 and 0.19 +/- 0.01 cm at the proximal and distal anastomoses. At explant, cells of mesenchymal origin (endothelial cells, smooth muscle cells, myofibroblasts) were embedded in the extracellular matrix of the graft scaffold. Minimal evidence of cellular inflammatory reaction and no aneurysmal dilatation or thrombus formation was detected. Variable degrees of hyperplasia were present at proximal and distal anastomoses. This preliminary study demonstrates that a collagen-based xenogeneic biomaterial provides a scaffold for cellularization when used for arterial reconstruction in dogs.