Massive bone reconstruction with heat‐treated bone graft loaded autologous bone marrow‐derived stromal cells and β‐tricalcium phosphate composites in canine models

Bone marrow‐derived stromal cells (BMSCs) contain mesenchymal stem cells that are capable of forming various mesenchymal tissues. We hypothesized that BMSCs and β‐tricalcium phosphate (β‐TCP) composites would promote the remodeling of large‐sized autologous devitalized bone grafts; therefore, the aim of this study was to evaluate the effects of the composites on the remodeling of autologous devitalized bone grafts. Autologous BMSCs cultured in culture medium containing dexamethasone (10^?7 M) were loaded into porous β‐TCP granules under low‐pressure. Theses BMSC/TCP composites were put into the bone marrow cavity of autologous heat‐treated bone (femoral diaphysis, 65‐mm long, 100°C, 30 min) and put back to the harvest site. In the contralateral side, β‐TCP without BMSC were used in the same manner as the opposite side as the control. Treatment with the BMSC/TCP composites resulted in a significant increase in thickness, bone mineral density, and matured bone volume of the cortical bone at the center of the graft compared to the control. Histological analysis showed matured regenerated bone in the BMSC loaded group. These results indicate that BMSC/TCP composites facilitated bone regeneration and maturation at the graft site of large‐sized devitalized bone. This method could potentially be applied for clinical use in the reconstruction of large bone defects such as those associated with bone tumors. © 2013 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 31:1308–1316, 2013

     

Prefabrication of a Vascularized Bone Graft With Beta Tricalcium Phosphate Using an In Vivo Bioreactor

We aimed to introduce an in vivo bioreactor‐vascular pedicle threaded through the central portion of a scaffold in which a vascularized bone graft was prefabricated using adenoviral human BMP‐2 gene (AdBMP2)‐modified bone marrow mesenchymal stem cells (BMSCs), beta tricalcium phosphate (β‐TCP), a vessel bundle, and muscularis membrane(group A). As controls, Adβgal‐BMSCs/β‐TCP granules, vessel bundle, and the muscularis membrane (group B); BMSCs/β‐TCP granules, vessel bundle, and muscularis membrane (group C); and β‐TCP granules, vessel bundle, and muscularis membrane (group D) were prepared. Formation of bone tissue and a vascular network was assessed by microangiography and histological methods 4 weeks after prefabrication. New cartilage and bone tissue in the space between β‐TCP granules (mainly endochondral bone) were confirmed by histology, and a de novo vascular network circulating from the vessel bundle through newly formed bone tissue was observed in group A. Formation of bone or cartilage was not observed in the control groups. We concluded that the in vivo bioreactor is a promising method for prefabrication of vascularized functional bone.     

Loss of wnt/β‐catenin signaling causes cell fate shift of preosteoblasts from osteoblasts to adipocytes

Wnt signaling is essential for osteogenesis and also functions as an adipogenic switch, but it is not known if interrupting wnt signaling via knockout of β‐catenin from osteoblasts would cause bone marrow adiposity. Here, we determined whether postnatal deletion of β‐catenin in preosteoblasts, through conditional cre expression driven by the osterix promoter, causes bone marrow adiposity. Postnatal disruption of β‐catenin in the preosteoblasts led to extensive bone marrow adiposity and low bone mass in adult mice. In cultured bone marrow–derived cells isolated from the knockout mice, adipogenic differentiation was dramatically increased, whereas osteogenic differentiation was significantly decreased. As myoblasts, in the absence of wnt/β‐catenin signaling, can be reprogrammed into the adipocyte lineage, we sought to determine whether the increased adipogenesis we observed partly resulted from a cell‐fate shift of preosteoblasts that had to express osterix (lineage‐committed early osteoblasts), from the osteoblastic to the adipocyte lineage. Using lineage tracing both in vivo and in vitro we showed that the loss of β‐catenin from preosteoblasts caused a cell‐fate shift of these cells from osteoblasts to adipocytes, a shift that may at least partly contribute to the bone marrow adiposity and low bone mass in the knockout mice. These novel findings indicate that wnt/β‐catenin signaling exerts control over the fate of lineage‐committed early osteoblasts, with respect to their differentiation into osteoblastic versus adipocytic populations in bone, and thus offers potential insight into the origin of bone marrow adiposity. © 2012 American Society for Bone and Mineral Research.     

Osteogenesis After Bone and Bone Marrow Transplantation: Studies of Cellular Behaviour Using Combined Myelo-osseous Grafts in the Subscorbutic Guinea Pig

In order to study factors influencing osteogenesis after bone and bone marrow transplantation, we have caused guinea pigs to become scorbutic, and looked at the cell morphology at sites of bone formation. We had previously studied normal guinea pigs and found that autologous marrow in intermuscular implants was associated with bone production by the ninth day, regardless of the type of stored allogeneic bone transplanted with it. in subscorbutic guinea pigs, using identical implants, bone did not appear within the first 13 days, and the cell population around the implants was different. These experiments support the dominant role of bone marrow cells in osteogenesis and cast further doubt on the primary role of devitalised bone as an inducer of bone formation. Interference with cell function by deprivation of a single essential molecule, Vitamin C, produces great change in the ability of cells to synthesise bone, or pre-osseous matrix.     

Osteogenesis after bone and bone marrow transplantation. Studies of cellular behaviour using combined myelo-osseous grafts in the subscorbutic guinea pig

In order to study factors influencing osteogenesis after bone and bone marrow transplantation, we have caused guinea pigs to become scorbutic, and looked at the cell morphology at sites of bone formation. We had previously studied normal guinea pigs and found that autologous marrow in intermuscular implants was associated with bone production by the ninth day, regardless of the type of stored allogeneic bone transplanted with it. In subscorbutic guinea pigs, using identical implants, bone did not appear within the first 13 days, and the cell population around the implants was different. These experiments support the dominant role of bone marrow cells in osteogenesis and cast further doubt on the primary role of devitalised bone as an inducer of bone formation. Interference with cell function by deprivation of a single essential molecule, Vitamin C, produces great change in the ability of cells to synthesise bone, or pre-osseous matrix.     

Bone ingrowth into porous calcium phosphate ceramics: influence of pulsing electromagnetic field

The effect of a pulsing electromagnetic field (PEMF) on bone ingrowth into porous hydroxyapatite (HA) and porous tricalcium phosphate (TCP) implanted in rabbit tibiae was studied. To quantitate the biological response, a recently developed method of surface measurement using a scanning electron microscope was used. The morphometrical findings in the HA pores demonstrated a significantly greater amount of bone and thicker bone trabeculae in the PEMF group as compared with the nonpulsed control group at 3 to 4 weeks postimplantation. No significant differences for these parameters were found in the TCP pores. Histologically, more bone and wider bone trabeculae were observed in the HA implants for the PEMF-treated animals at the early time periods when compared with those of the control animals. Alternatively, the histological findings of the TCP implants were similar between these two groups. These histological results tended to correlate with the morphometrical data. Together, these results suggest that accelerated bone formation and bone maturation occurred in response to PEMF in the HA pores but was without effect in the TCP pores. This stimulatory effect is most significant after 3-4 weeks of PEMF stimulation.     

Skeletal repair in rabbits using a novel biomimetic composite based on adipose‐derived stem cells encapsulated in collagen I gel with PLGA‐β‐TCP scaffold

In bone tissue engineering, the cell distribution mode in the scaffold may affect in vivo osteogenesis. Therefore, we fabricated a novel biomimetic construct based on a combination of rabbit adipose‐derived stem cells (rASCs) encapsulated in collagen I gel with a PLGA‐β‐TCP scaffold (rASCs‐COL/PLGA‐β‐TCP, group A), the combination of rASCs and PLGA‐β‐TCP (rASCs/PLGA‐β‐TCP, group B), the combination of collagen I gel and PLGA‐β‐TCP (COL/PLGA‐β‐TCP, group C), and PLGA‐β‐TCP scaffold (group D). The composites were implanted into a 15‐mm length critical‐sized segmental radial defect. The results were assessed by histology, radiographs, bone mineral density (BMD), and mechanical testing. After 24 weeks, the medullary cavity recanalized, bone was rebuilt, and molding finished, the bone contour remodeled smoothly and the scaffold degraded completely in group A. The BMDs and mechanical properties were similar to normal. However, the bone defect remained unrepaired in groups B, C, and D. Moreover, the scaffold degradation rate in group A was significantly higher than the other groups. Thus, enhanced in vivo osteogenesis of rASCs wrapped in collagen I gel combined with PLGA‐β‐TCP was achieved, and the bone defect was repaired. We hope this study provides new insights into ASCs‐based bone tissue engineering. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:252–257, 2010     

Novel application of HA‐TCP biomaterials in distraction osteogenesis shortened the lengthening time and promoted bone consolidation

This study tested the hypothesis that use of biomaterials in distraction osteogenesis (DO) would reduce the treatment time and enhance bone formation quality. A 1.0‐cm tibial shaft was removed in the left tibia of 36 rabbits. Rabbits were randomly divided into three groups: group A, the defect gap was reduced with the tibia shortened for 1.0‐cm; group B, the defect gap was filled with 1.0‐cm restorable porous hydroxyapatite and Tri‐calcium phosphates cylindrical block (HA/TCP block, diameter is 0.5‐cm); group C, The 1.0‐cm defect gap was reduced 0.5 cm and the remaining 0.5‐cm defect gap was filled with the 0.5‐cm HA/TCP block. The tibia was then fixed with unilateral lengthener; for groups A and C; lengthening started 7 days after surgery at a rate of 1.0 mm/day, in two steps. Group A received lengthening for 10 days and group C for 5 days, there was no lengthening for group B. All animals were terminated at day 37 following surgery. The excised bone specimens were subject to microcomputed tomography (micro‐CT), mechanical testing, and histological examinations. Bone mineral density and content and tissue mineral density and content, as well as the mechanical properties of the regenerates were significantly higher in group C compared to groups A and B. Micro‐CT and histological examinations also confirmed that the regenerates in Group C had most advanced bone formation, consolidation, and remodeling compared to other groups. In conclusion, the combined use of biomaterials and DO technique can reduce the treatment time and enhance bone consolidation in bone defect management. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27: 477–482, 2009     

骨髓基质干细胞联合血管束植入构筑血管化组织工程骨修复兔大段骨缺损

目的 研究兔骨髓基质干细胞(BMSCs)联合动静脉血管束植入异种脱蛋白松质骨(XDCB)构筑血管化组织工程骨修复兔桡骨中远段完全骨膜骨缺损的能力. 方法 从兔髂嵴捕骨髓培养制备兔BMSCs,将第5代BMSCs种植于多孔XDCB,并进行成骨诱导2周制备组织工程骨,手术中分离兔桡动、静脉血管束.动物模型为制备24只兔舣侧桡骨中远段完全骨膜骨缺损1.5 cm共48侧,分4组修复(n=12),A组为空白未治疗组,B组为单纯材料+血管束植入组(XDCB+VB),C组为组织工程骨组(XDCB+BMSCs),D组为组织工程骨+血管束植入组(XDCB+BMSCs+VB),符组交叉配对.分别于术后4、8、12周行X线片、大体解剖、组织切片、生物力学等检查,观察各组骨缺损修复效能及移植物血管化情况.结果 D组骨缺损修复效能(术后12周新骨面积比2.02%±0.16%)及血管化情况(术后12周血管面积比6.89%±0.32%)优于C组(1.50%±0.28%和3.17%±0.19%),而C组又优于B组(1.59%±0.19%和6.52%±0.23%),A组骨缺损未修复,各组结果差异有统计学意义(P<0.05).结论 BMSCs联合动静脉血管束植入构筑的血管化组织工程骨能促进成骨过程和新生骨的血管化,显著提高组织工程骨修复大段骨缺损的能力.     

Heterotopic osteogenesis in porous ceramics induced by marrow cells

When untreated porous calcium phosphate ceramics were transplanted into subcutaneous (s.c.) or intramuscular (i.m.) sites, fibrovascular tissue grew in the pore region without evidence of bone formation. However, when these same ceramics were combined with syngeneic marrow cells, osteogenesis was observed inside the pore region of the implanted ceramic. The osteogenesis began on the surface of the pore region at approximately 3 weeks postimplantation by a process of intramembranous bone formation, with the de novo bone tissue observed directly interfacing with the ceramic surface. Infrequently, small isolated areas showed cartilage formation with no noticeable endochondral ossification. At 4 weeks postimplantation of the ceramic with marrow cells, the osteogenesis in the ceramic accompanied an observed increase in compressive strength, rigidity, and energy absorption of the ceramic. These results suggest that a combination of porous ceramics and marrow cells may be useful for clinical problems requiring osseous reconstruction.     

羟基磷灰石材料结合牵拉成骨技术修复骨缺损的实验研究

 目的 探讨羟基磷灰石（hydroxyapatite/tri-calcium phosphase,HA/TCP）材料结合牵拉成骨技术是否能减少治疗骨缺损所需的时间并促进骨的矿化过程。方法 将36只健康雄性新西兰大白兔（20~24周,体重2.2~2.8 kg）随机分为3组,每组12只。于左胫腓关节下方做1 cm的胫骨缺损。A组：将骨缺损两端靠拢,外固定架加压固定。B组：将1 cm长的HA/TCP材料填入1 cm的骨缺损中,外固定架加压固定。C组：将骨缺损两端拉近至间距0.5 cm,将0.5 cm长的HA/TCP材料填入剩余的0.5 cm骨缺损中,外固定架加压固定。除B组外,A、C组均于术后7 d开始延长胫骨,A组延长10 d,C组延长5 d。在术后即刻、12、17、27、37 d用“C”型臂X线机观察各组新生骨的矿化过程。所有动物于术后37 d处死,取新生骨样本测量骨矿物质含量（BMC）和组织矿物质含量（TMC）,行胫骨扭转实验及组织病理学分析。结果 C组BMC为（454.44±89.98） mg和TMC为（454.40±89.97） mg显著高于A、B两组。C组新生骨的最大扭矩、新生骨的成熟度均明显高于A、B两组,骨的矿化和重塑也较快。C组在观察期总共37 d内骨愈合良好,但A、B两组在37 d内并未达到骨愈合。结论 在兔胫骨缺损延长模型中,与单一疗法相比,联合应用羟基磷灰石材料和牵拉成骨技术可减少治疗时间并促进骨的矿化。     

Promotion of osteogenesis in tissue‐engineered bone by pre‐seeding endothelial progenitor cells‐derived endothelial cells

In addition to a biocompatible scaffold and an osteogenic cell population, tissue‐engineered bone requires an appropriate vascular bed to overcome the obstacle of nutrient and oxygen transport in the 3D structure. We hypothesized that the addition of endothelial cells (ECs) may improve osteogenesis and prevent necrosis of engineered bone via effective neovascularization. Osteoblasts and ECs were differentiated from bone marrow of BALB/c mice, and their phenotypes were confirmed prior to implantation. Cylindrical porous polycaprolactone (PCL)‐hydroxyapatite (HA) scaffolds were synthesized. ECs were seeded on scaffolds followed by seeding of osteoblasts in the EC‐OB group. In the OB group, scaffolds were only seeded with osteoblasts. The cell‐free scaffolds were denoted as control group. A 0.4‐cm‐long segmental femur defect was established and replaced with the grafts. The grafts were evaluated histologically at 6 weeks postimplantation. In comparison with the OB group, the EC‐OB group resulted in a widely distributed capillary network, osteoid generated by osteoblasts and absent ischemic necroses. Pre‐seeding scaffold with ECs effectively promoted neovascularization in grafts, prevented the ischemic necrosis, and improved osteogenesis. The integration of bone marrow‐derived ECs and osteoblasts in porous scaffold is a useful strategy to achieve engineered bone. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:1147–1152, 2008     

Customized,degradable, functionally graded scaffold for potential treatment of early stage osteonecrosis of the femoral head

Osteonecrosis of the femoral head (ONFH) is a debilitating disease that results in progressive collapse of the femoral head and subsequent degenerative arthritis. Few treatments provide both sufficient mechanical support and biological cues for regeneration of bone and vascularity when the femoral head is still round and therefore salvageable. We designed and 3D printed a functionally graded scaffold (FGS) made of polycaprolactone (PCL) and β‐tricalcium phosphate (β‐TCP) with spatially controlled porosity, degradation, and mechanical strength properties to reconstruct necrotic bone tissue in the femoral head. The FGS was designed to have low porosity segments (15% in proximal and distal segments) and a high porosity segment (60% in middle segment) according to the desired mechanical and osteoconductive properties at each specific site after implantation into the femoral head. The FGS was inserted into a bone tunnel drilled in rabbit femoral neck and head, and at 8 weeks after implantation, the tissue formation as well as scaffold degradation was analyzed. Micro‐CT analysis demonstrated that the FGS‐filled group had a significantly higher bone ingrowth ratio compared to the empty‐tunnel group, and the difference was higher at the distal low porosity segments. The in vivo degradation rate of the scaffold was higher in the proximal and distal segments than in the middle segment. Histological analysis of both non‐decalcified and calcified samples clearly indicated new bone ingrowth and bone marrow‐containing bone formation across the FGS. A 3D printed PCL‐β‐TCP FGS appears to be a promising customized resorbable load‐bearing implant for treatment of early stage ONFH. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1002–1011, 2018.

     

Autogeneic bone marrow and porous biphasic calcium phosphate ceramic for segmental bone defects in the canine ulna.

The purpose of this study was to evaluate a porous biphasic hydroxyapatite-calcium phosphate ceramic as a modifier and extender of an autogeneic marrow graft for filling a 2.5-cm segmental bony defect. Twenty adult mongrel dogs were surgically treated to create diaphyseal defects in the left ulnae. The defects were (1) filled with autogeneic bone marrow mixed with granular hydroxyapatite-tricalcium phosphate ceramic (granular ceramic); (2) grafted with a solid block of ceramic soaked in autogeneic bone marrow (block ceramic); (3) received no graft (no implant); or (4) were grafted with autogeneic bone marrow alone (bone marrow). All animals were followed clinically and roentgenographically for 24 weeks and then killed. Repair of diaphyseal defects with the block ceramic led to three solid unions and three fibrous unions; with the granular ceramic implants and marrow, the defects of five dogs formed solid unions, and one progressed to a fibrous union. Defects in all five dogs grafted with autogeneic bone marrow united. The three dogs with no implant formed nonunions. Histology showed normal marrow and only a light immune reaction. Complete bridging of the defect in the dogs treated with the granular ceramic occurred significantly earlier than bridging in the dogs grafted with bone marrow alone. Histomorphometry, performed on the block ceramic implants indicated active resorption of ceramic. Clinically, addition of ceramic to a marrow graft improved the handling characteristics of the graft material and accelerated healing according to roentgenographic evaluation.     

Enhanced Bone Bonding of the Hydroxyapatite/β‐Tricalcium Phosphate Composite by Electrical Polarization in Rabbit Long Bone

A review of the osteogenic cell activity and new bone growth in the regions bordering negatively charged surfaces of polarized Hydroxyapatite/β‐tricalcium phosphate (HA/TCP) composites implanted in the long bone in rabbits was conducted. Polarized and non‐polarized HA/TCP specimens were implanted into the right and left femoral condyle, respectively (each n = 10). After 3 and 6 weeks, five rabbits were sacrificed in each group, and histological analysis was administered. Large cuboidal‐shaped osteoblastic cells were predominantly observed lining the newly formed bone on the negatively charged surface (N‐surface) in the polarized HA/TCP implants. The TRAP‐positive multinucleated cells were observed extensively in the newly formed bone on the N‐surfaces compared with the 0‐surface and adhered directly to the HA/TCP composite. The bone area (B.Ar) value, newly formed bone area contacting the implant, and contact length (C.Le) value, percentage length of newly formed bone directly attaching to the implant, on both the 0‐ and N‐surface increased significantly with time in each group. Both the B.Ar and C.Le value on the N‐surface were significantly greater than those on the 0‐surface after 3 and 6 weeks. The number of TRAP‐positive cells/total length value on the N‐surface was significantly greater than that on the 0‐surface after 3 and 6 weeks postoperatively. It is hypothesized that electrical charge acquired by electrical polarization treatment may modify the biochemical and biophysical processes of the osteogenic cells, resulting in enhanced new bone formation and direct bonding between the recipient bone and implants.     

Exogenous phytoestrogenic molecule icaritin incorporated into a porous scaffold for enhancing bone defect repair

This study was designed to develop a bioactive scaffold to enhance bone defect repair in steroid‐associated osteonecrosis (SAON). Icaritin, a metabolite of the herb Epimedium, has been identified as an angiogenic and osteogenic phytomolecule. Icaritin was homogenized into poly lactic‐co‐glycolic acid/tricalcium phosphate (PLGA/TCP) to form an icaritin‐releasing porous composite scaffold (PLGA/TCP/icaritin) by fine‐spinning technology. In vitro, high performance liquid chromatography was used to determine the release of icaritin during degradation of PLGA/TCP/icaritin. The osteogenic effects of PLGA/TCP/icaritin were evaluated using rat bone marrow mesenchymal stem cells (BMSCs). In vivo, the osteogenic effect of PLGA/TCP/icaritin was determined within a bone tunnel after core decompression in SAON rabbits and angiography within scaffolds was examined in rabbit muscle pouch model. In vitro study confirmed the sustainable release of icaritin from PLGA/TCP/icaritin with the bioactive scaffold promoting the proliferation and osteoblastic differentiation of rat BMSCs. In vivo study showed that PLGA/TCP/icaritin significantly promoted new bone formation within the bone defect after core decompression in SAON rabbits and enhanced neovascularization in the rabbit muscle pouch experiment. In conclusion, PLGA/TCP/icaritin is an innovative local delivery system that demonstrates sustainable release of osteogenic phytomolecule icaritin enhancing bone repair in an SAON rabbit model. The supplement of scaffold materials with bioactive phytomolecule(s) might improve treatment efficiency in challenging orthopedic conditions. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31:164–172, 2012     

Promotion of normal healing of bone defects under estrogen deficiency by implantation of beta‐tricalcium phosphate composed of rod‐shaped particles

We compared the healing of bone defects in ovariectomized rats implanted with beta‐tricalcium phosphate (β‐TCP) composed of rod‐shaped particles, which were prepared using the applied hydrothermal method (HTCP), and that of bone defects implanted with conventional β‐TCP composed of globular‐shaped particles (CTCP), which were prepared by normal sintering. Eight‐week‐old female Wistar rats were ovariectomized, and 2 weeks after the operation, 0.5‐ to 0.6‐mm diameter spherical granules of each ceramic were implanted in a bone defect created in the distal end of the femur. Four, 8, and 12 weeks after implantation, the amount of newly formed bone implanted with HTCP was significantly larger than that implanted with CTCP and was equivalent to that in non‐ovariectomized sham‐operated rats. Without implantation, spontaneous repair of the trabecular bone was barely observed. The physiological structure of the trabecular network was maintained in the region implanted with HTCP, but that in the region implanted with CTCP was severely destroyed. Gene expression microarray analysis revealed that the expression of genes involved in interferon signaling pathways was upregulated in osteoclasts cultured on HTCP compared with that cultured on CTCP. Our results suggest that the microstructure of β‐TCP affected the biological behavior of osteoclasts and regulated local bone metabolism. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:189–196, 2014.     

Plasticizer di(2‐ethylhexyl)phthalate interferes with osteoblastogenesis and adipogenesis in a mouse model

Plasticizer di(2‐ethylhexyl)phthalate (DEHP) can leach from medical devices such as blood storage bags and the tubing. Recently, epidemiological studies showed that phthalate metabolites levels in the urine are associated with low bone mineral density (BMD) in older women. The detailed effect and mechanism of DEHP on osteoblastogenesis and adipogenesis, and bone loss remain to be clarified. Here, we investigated the effect and mechanism of DEHP and its active metabolite mono(2‐ethylhexyl)phthalate (MEHP) on osteoblastogenesis and adipogenesis. The in vitro study showed that osteoblast differentiation of bone marrow stromal cells (BMSCs) was significantly and dose‐dependently decreased by DEHP and MEHP (10–100 µM) without cytotoxicity to BMSCs. The mRNA expressions of alkaline phosphatase, Runx2, osteocalcin (OCN), Wnt1, and β‐catenin were significantly decreased in DEHP‐ and MEHP‐treated BMSCs during differentiation. MEHP, but not DEHP, significantly increased the adipocyte differentiation of BMSCs and PPARγ mRNA expression. Both DEHP and MEHP significantly increased the ratios of phosphorylated β‐catenin/β‐catenin and inhibited osteoblastogenesis, which could be reversed by Wnt activator lithium chloride and PPARγ inhibitor T0070907. Moreover, exposure of mice to DEHP (1, 10, and 100 mg/kg) for 8 weeks altered BMD and microstructure. In BMSCs isolated from DEHP‐treated mice, osteoblastogenesis and Runx2, Wnt1, and β‐catenin expression were decreased, but adipogenesis and PPARγ expression were increased. These findings suggest that DEHP and its metabolite MEHP exposure may inhibit osteoblastogenesis and promote adipogenesis of BMSCs through the Wnt/β‐catenin‐regulated and thus triggering bone loss. PPARγ signaling may play an important role in MEHP‐ and DEHP‐induced suppression of osteogenesis. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1124–1134, 2018.

     

Demineralized bone implants

There are three mechanisms of bone formation that underlie the use of the different types of implants. In osteogenesis, viable osteoblasts and preosteoblasts are transplanted from one part of the body to the site where new bone is needed; cancellous marrow grafts are an example of such osteogenic engraftment. In osteoconduction, the implant does not provide many viable cells but rather acts as a scaffolding for the ingrowth of new bone from the margins of the defect with the concurrent resorption of the implant; cortical bone grafts or banked bone segments are examples of this "creeping substitution." In osteoinduction, the implant stimulates the transformation of connective tissue to produce endochondral bone, even in extraskeletal sites; demineralized bone implants promote bone formation by osteoinduction. The physiology, cell biology, biochemistry, and endocrinologic regulation of induced osteogenesis are areas of active investigation. Fresh autogenous cancellous bone grafts are preferred for non-stress-bearing defects, but are often of limited availability for extensive procedures, especially in infants. Demineralized bone implants have been used successfully in certain types of craniomaxillofacial, orthopedic, periodontal, and hand reconstruction. Tissue transformation may become as important to reconstructive surgery as is tissue transplantation.     

新型仿生活性人工骨诱导兔腰椎横突间脊柱融合研究

目的采用先进快速成形技术（RP）结合骨组织工程方法研制新型仿生活性人工骨，并探讨其在兔腰椎横突间脊柱融合的应用情况。方法首先采用RP制备薄块型聚乳酸一聚羟乙酸／磷酸三钙（PLGA／TCP）人工骨载体，进而高效复合牛骨形态发生蛋白（bBMP）以制备仿生活性人工骨。扫描电镜观察载体材料及人工骨超微结构。将健康新西兰兔28只（平均体重4.1kg）随机均分为A、B两组（每组14只）。A组：于兔腰4～5右、左侧横突间分别植入仿生活性人工骨（A1组）、自体髂骨（A2组）；B组：于兔腰4～5右、左侧横突间分别植入复合自体新鲜红骨髓的PLGA／TCP载体材料（B1组）、单纯PLGA／TCP载体材料（B2组）。于术后6周和12周，定期大体观察、手法检测、组织学[苏木素-伊红（HE）、三色法及四环素-钙黄绿素荧光检测]和影像学方法（X、CT）系统评价脊柱融合情况。结果RP制备的PLGA／TCP载体具有规则的空间支架结构、相互贯通的孔隙及材料表面微孔特征。这些均有利于bBMP的高效复合。动物实验结果显示，A1组仿生活性人工骨植入具有强的诱骨活性及骨性融合能力，不仅成骨早、新骨形成量大，而且在新骨形成及改塑的同时载体材料逐渐降解。术后12周可形成较为典型的骨小梁及骨髓结构，骨代谢活性亦接近正常。A2组自体髂骨移植能达到良好骨性融合，但术后12周所形成的新骨结构尚须进一步塑形及完善。B1组、B2组术后12周仍遗有较多的载体材料有待降解，基本无成骨能力。术后12周，A1、A2、B1、1324组横突间融合率分别为100．0％、58．3％、18．2％和0％，A1组融合率最高（P〈0．01）。结论先进RP制备的PLGA／TCP载体不仅具有良好的空间超微结构及孔隙特性，而且能高效复合bBMP以正确构建新型组织工程人工骨。该仿生活性人工骨诱导兔横突间脊柱融合获得成功．为生物制造脊柱外科所需的新型、高效人工骨移植材料奠定了重要基础。