脂肪组织来源干细胞复合胶原构建组织工程皮肤修复皮肤缺损

目的:探讨脂肪组织来源干细胞(ADSC)作为皮肤组织工程种子细胞修复皮肤缺损的可能性。方法:采用胶原酶消化SD大鼠脂肪组织,经体外扩增后接种于胶原凝胶构建组织工程活性皮肤替代物。15只SD大鼠随机分成3组:ADSC复合胶原组织工程活性皮肤替代物实验组、单纯胶原移植组和空白对照组。将各组相应材料分别移植于SD大鼠背部皮肤缺损处,比较观察创面修复效果。结果:ADSC复合胶原实验组、单纯胶原移植组和空白对照组创面愈合时间分别为:(14.3±1.7),(16.9±2.5)和(21.2±4.2)d,差异有显著性意义(P<0.05)。组织学观察显示,ADSC复合胶原实验组成纤维细胞、微血管、胶原纤维均较单纯胶原移植组和空白对照组明显增多。结论:ADSC复合胶原构建的组织工程活性皮肤替代物移植后可加速新生血管形成、促进创面修复。提示以ADSCs作为种子细胞复合胶原构建的组织工程皮肤可用于皮肤缺损的修复治疗。     

CGF与脂肪干细胞联合应用修复大鼠全层皮肤缺损的研究

目的:探讨以明胶海绵为支架材料,复合浓缩生长因子(concentrated growth factor,CGF)诱导的大鼠脂肪干细胞(adipose-derived stem cells,ADSCs)得到的组织工程皮肤替代物,对大鼠全层皮肤缺损愈合的作用。方法:取健康雄性大鼠脂肪组织分离、培养并鉴定脂肪干细胞,观察CGF对其增殖、分化的影响。在健康雌性大鼠背部制造一直径3 cm全层皮肤缺损,其上覆盖复合脂肪干细胞的明胶海绵(A组)、无细胞的明胶海绵(B组)或不作处理单纯包扎(C组)。术后观察各组缺损的愈合情况,并于7、14、21 d及完全愈合后,取背部全层皮肤作组织学观察。结果:CGF能促进体外培养的ADSCs增殖;术后伤口渗出物、红肿等炎症反应A组相似文献   

复合ADSCs/β-TCP组织工程骨与PRF修复下颌骨缺损的实验研究

目的:探讨脂肪干细胞/β-磷酸三钙(ADSCs/β-TCP)组织工程骨复合富血小板纤维蛋白(PRF)在修复下颌骨缺损中的作用。方法:新西兰大白兔27只,采用随机对照动物实验,随机分为3组,每组9只,培养家兔自体ADSCs,制备自体PRF。在每只兔子的双侧下颌骨制备下颌骨缺损模型。A组:将单纯的β-磷酸三钙(β-TCP)植入双侧下颌骨缺损区,B组:将ADSCs/β-TCP植入双侧下颌骨缺损区,C组:将ADSCs/β-TCP与PRF植入双侧下颌骨缺损区。分别于术后2、4、8周分别处死9只大白兔,并通过大体观察、X线片、以及组织切片观察下颌骨缺损的修复情况。结果:C组在2、4、8周3个时间点的成骨情况明显优于其他2组(P<0.01)。A组与B组之间的成骨情况差异无显著性(P>0.05)。结论:ADSCs/β-TCP复合物与PRF联合应用可促进下颌骨缺损的修复。     

PRF促进小鼠全层皮肤损伤修复的实验研究

目的:探讨富血小板纤维蛋白(PRF)对小鼠皮肤损伤的修复作用。方法:采用8~10周的Balb/C小鼠21只,将其分为3组,分别为皮肤缺损空白对照组,上市产品对照组(AD甲壳素),冻干异种PRF颗粒组,在背部制备1个1.5 cm圆形全层皮肤损伤创面。通过大体观察,测定伤口愈合时间和愈合率,并在第21天,取皮肤组织做病理切片,观察组织愈合情况。结果:伤后21 d,各组创面逐渐缩小,冻干异种PRF颗粒组创面面积明显小于其余各组(P＜0.01),并且愈合时间较其余各组短。结论:PRF可以明显促进创面愈合速度,提高愈合质量,为临床皮肤损伤修复提供新的治疗方法。     

脂肪干细胞复合成骨性干细胞膜片用于修复兔颅骨缺损的实验研究

目的　细胞膜片技术已经被证实了可以成功用于多种组织再生,我们前期实验证明了脂肪干细胞（ADSCs）可以显著促进成骨性细胞膜片（BMSCs sheet）的异位成骨能力。因此,本实验的目的是验证ADSCs能否促进成骨性BMSCs膜片的骨缺损修复能力。方法：ADSCs和BMSCs分别取自同一个供体的腹股沟和髂骨骨松质,通过连续成骨诱导的方法制备成骨性BMSCs膜片,并分析BMSCs膜片的特性;然后将ADSCs复合在BMSCs膜片上,构架ADSCs-BMSCs膜片复合体,移植到兔颅骨缺损模型,单纯BMSCs膜片和颅骨缺损组作为对照。移植八周后,Micro-CT扫描和组织学分析其骨修复再生情况。结果：BMSCs膜片由含有大量矿化结节的多层细胞膜片构成;移植八周后,相比于对照组,实验组新生组织骨密度更高,新生矿化组织量也更多,同时新生骨组织/总新生组织体积（BV/TV）的比值也更高,表明ADSCs显著提高了BMSCs膜片的骨缺损修复能力。结论：ADSCs复合BMSCs膜片可以明显促进兔颅骨缺损的骨再生修复,该方法为促进骨修复重建提供了一种新的方法。     

纳米羟基磷灰石复合胶原材料负载骨髓基质干细胞修复颅骨极限缺损的实验研究

目的:探讨纳米羟基磷灰石复合胶原材料(nHAC)负载骨髓基质干细胞作为组织工程骨在修复颅骨极限缺损中的作用。方法:20只新西兰大白兔随机分为复合细胞组、单纯材料组、空白对照组。复合细胞组取自体骨髓基质干细胞并扩增诱导培养后通过负压抽吸及细胞基质膜包裹将细胞与材料复合。材料植入颅骨缺损后8周取材,行大体、X线、组织学观察及灰量测量来研究它们的骨修复能力。结果:复合细胞组成骨能力最强,且材料边缘及内部均有较好的成骨,单纯材料组有部分成骨,空白对照组未修复骨缺损。结论:纳米羟基磷灰石复合胶原材料负载骨髓基质干细胞后是一种较为理想的骨替代材料。     

脂肪来源干细胞膜片与支持材料结合能力的初步探讨

目的:探讨大鼠脂肪来源干细胞体外构建细胞膜片方法,并分析其同牙本质磨片和CBB(ceramic bovine bone,CBB)颗粒的结合能力和增殖影响。方法:从出生7 d大鼠体内提取脂肪组织并获得纯化的脂肪来源干细胞(ADSCs,adipose-derived mesenchymal stem cell), 采用四唑盐(MTT)比色法检测ADSCs增殖能力的变化;ADSCs经多向诱导分化后检测其向成骨细胞、脂肪细胞、成软骨细胞分化能力;ADSCs体外构建细胞膜片后,电镜下观察ADSCs膜片与CBB颗粒和牙本质磨片的结合能力,MTT比色法检测ADSCs和ADSCs膜片在材料上的增殖能力。结果:纯化的ADSCs具有良好的增殖能力,经体外诱导培养后向成骨细胞、脂肪细胞、成软骨细胞分化,电镜结果表明ADSCs膜片与牙本质磨片结合能力优于CBB颗粒,ADSCs膜片在牙本质磨片上的增殖能力优于CBB颗粒。结论:体外培养的大鼠脂肪来源干细胞具有种子细胞的特性,体外构建细胞膜片后与牙本质磨片的结合能力和增殖能力高于CBB颗粒,为细胞膜片治疗牙周缺损奠定基础。     

脂肪干细胞在牙周组织工程学中的的应用前景

脂肪干细胞(adipose tissue-derived stem cells,ADSCs),是一种新的组织工程的干细胞来源.本文阐述了脂肪组织干细胞优势,并对脂肪干细胞的培养与鉴定以及成骨分化潜能,在牙周组织工程学中的潜在应用前景作一综述.     

脂肪干细胞复合载淫羊藿苷支架材料对兔下颌骨缺损的修复

目的:研究脂肪干细胞(ADSCs)作为种子细胞复合淫羊藿苷支架材料修复兔下颌骨缺损的效果.方法:体外培养ADSCs,茜素红染色检测骨向分化.实验组在骨缺损处植入ADSCs复合载淫羊藿苷支架材料;对照组2植入单纯纳米羟基磷灰石(n-HA)材料;对照组2植入淫羊藿苷/n-HA复合材料;空白组不植入任何材料.应用X线、组织学、扫描电镜观察并比较植入物与骨组织交界处的成骨修复情况.结果:实验组骨缺损区完全被骨组织修复,边界已融合;对照组1部分修复,有少量炎性渗出;对照组2大部分已修复,边界模糊;空白组基本未见修复.结论:ADSCs复合载淫羊藿苷支架材料具有骨缺损修复能力,其效果优于淫羊藿苷/n-HA材料及单纯n-HA材料.     

应用牙周膜干细胞—牙囊干细胞复合细胞膜片同种异体移植修复比格犬牙周组织缺损的研究

目的 拟构建比格犬牙周缺损模型,观察牙周膜干细胞—牙囊干细胞复合细胞膜片修复牙周组织缺损的效果.方法 分离培养比格犬牙囊干细胞、牙周膜干细胞,制备牙囊干细胞细胞膜片、牙周膜干细胞细胞膜片以及以上2种细胞的复合细胞膜片,将膜片混合支架材料同种异体植入成年雄性健康比格犬牙周缺损模型,对照组只植入支架材料.分别于移植后1周、2周、4周、6周、8周、12周检测探诊深度、牙龈退缩、附着丧失情况.12周后观察分析牙周再生的组织形态学变化.结果 牙周临床检查及组织学形态学分析显示,3个膜片移植组牙周再生效果较对照组效果明显,其中复合细胞膜片移植组明显高于其余两个单一膜片移植组(P<0.05).结论 牙周膜干细胞—牙囊干细胞复合膜片同种异体移植更有利于修复比格犬牙周缺损.     

Human deciduous teeth dental pulp cells with basic fibroblast growth factor enhance wound healing of skin defect

In this research, we examined the effect on wound healing applying basic fibroblast growth factor (b-FGF) that is approved for clinical use to enhance wound healing and human deciduous teeth dental pulp cells (hDPCs) in clinics, but that have been attracting attention as a novel stem cell source in recent years. Human deciduous teeth were harvested from healthy volunteers, and hDPCs were isolated. We used a nude mouse full-thickness skin defect model and evaluated wound healing by macroscopic view and histologic and histomorphometric analysis. The mice were randomly divided into 4 groups: phosphate-buffered saline-treated group (control group), b-FGF-treated group (b-FGF group), hDPC-treated group (hDPC group), and hDPC and b-FGF-treated group (hDPC/b-FGF group). Basic fibroblast growth factor and hDPC groups accelerated wound healing compared with the control group. There was no statistically significant difference in wound healing observed between the hDPC and b-FGF groups. The hDPC/b-FGF group demonstrated accelerated wound healing compared with other groups. At day 14, PKH26-positive cells were surrounded by human type I collagen in hDPC and hDPC/b-FGF groups in immunohistologic evaluation. Significantly increased collagen fibril areas in wound tissues were observed in b-FGF, hDPC, and hDPC/b-FGF groups as compared with the control group at days 7 and 14. Our results showed that the hDPC/b-FGF group significantly promotes wound healing compared with other groups. This study implies that deciduous teeth that are currently considered as medical spare parts might offer a unique stem cell resource for potential of new cell therapies for wound healing in combination with b-FGF.     

Effect of low-dose electron radiation on rat skin wound healing

The aim of this study was to assess the effect of low-dose electron irradiation on morphological features of the wound healing process in rat skin. Surgical wounds were inflicted with a 2.3 x 1.4 cm template on 84 male rats that were assigned to 4 groups: IG1, immediately irradiated; IG2, irradiated 3 days after inflicting the wound; CG1 and CG2, control groups. Rats in IG1 and IG2 groups had their wounds exposed to 1 Gy of 6 MeV electron beam radiation, immediately after surgery and on the third postoperative day, respectively. Qualitative and histophotometric evaluations of tissue repair structures were carried out. Data were analyzed by ANOVA and Tukey's test (alpha = 0.05) and regression analysis. The repair process was delayed since the first sacrifice time in both irradiated groups, but in IG1, wound healing was closer to that of CG1; whereas in IG2, the delay was more pronounced. Based on the histological findings, it is possible to conclude that a low-dose of electron radiation delayed tissue repair in rat skin. The delay was longer in the skin irradiated 3 days after the beginning of tissue repair. However, the low-energy electron irradiation did not prevent wound healing.     

In vivo alveolar bone regeneration by bone marrow stem cells/fibrin glue composition

The repair of alveolar bone defects caused by trauma, periodontal diseases and inflammation is still a challenge for both researchers and clinicians. Although there are many attempts to regenerate bone based on different seed cells and scaffolds, the results are still unsatisfactory. This study aims to clarify whether it could be efficient to reconstruct the alveolar bone by the combination of bone marrow stem cells (BMSCs) without pre-osteoinduction in vitro with fibrin glue (FG). The BMSCs were obtained from 2-week-old Sprague-Dawley (SD) rats and expanded in vitro with non-introduction. Afterwards, they were composited with FG for in vivo implantation. The animal models of traumatic alveolar bone defects were established bilaterally in the maxilla of 15 rats which were randomly divided into 3 groups. The BMSCs/FG composition was transplanted into 5 rats of the treated group. Another 5 rats in the negative control group were transplanted by pure FG without BMSCs. The rest 5 rats served as the blank control. Gross observation and histological analysis were made to evaluate the new bone formation 6 weeks after transplantation. Micro-CT was also used to estimate the bone healing through three-dimensional reconstruction and the bone density analysis. The amount of new bone formed in the treated group was significantly greater than the negative and blank control. Our results suggest that the strategy of combing BMSCs with FG is effective in the repair of alveolar bone defects. Its clinical application is promising.     

富血小板纤维蛋白对兔脂肪干细胞成骨分化的影响

目的：体外分离培养兔脂肪干细胞（adipose—derived stem cells,ADSCs）,鉴定其分化能力并观察富血小板纤维蛋白（platelet—rich fibrin,PRF）对ADSCs成骨分化的影响。方法：取新西兰兔腹股沟处的脂肪组织,将其分离获得脂肪干细胞,培养至第三代用于实验。分别以油红O、茜素红染色鉴定其成脂和成骨分化能力。取兔耳中央动脉血一次离心法制备PRF膜。将脂肪干细胞分为2组：对照组不含PRF膜;实验组含PRF膜。用碱性磷酸酶（ALP）试剂盒检测不同时间点PRF对ADSCs成骨分化的影响。结果：脂肪干细胞呈长梭形贴壁生长;油红O及茜素红染色均呈阳性;在不同的时间点,实验组ALP活性值均高于对照组（P〈0．05）。结论：ADSCs具有成骨的潜能,且PRF可以促进ADSCs向成骨细胞分化。     

The effect of basic fibroblast growth factor on delayed-replanted monkey teeth

BACKGROUND: Basic fibroblast growth factor (bFGF; FGF-2) has been reported to facilitate wound healing and periodontal regeneration in experimental alveolar bone defects. The purpose of this study was to evaluate histologically the effect of topically applied bFGF with or without fibrin glue on delayed-replanted monkey teeth prone to replacement resorption. METHODS: Forty-five roots from five monkeys were endodontically treated aseptically and then extracted as atraumatically as possible. Ten negative control roots were replanted immediately, while 12 positive control roots were allowed to bench dry for 1 hour prior to replantation, both without further treatment. Roots in the two experimental groups were bench dried for 1 hour, rinsed with saline, and then replanted into sockets filled with bFGF with (11 roots) or without (12 roots) fibrin glue. After 12 weeks, histological sections were prepared and evaluated according to morphometric analysis as complete healing or unfavorable healing composed of inflammatory resorption and replacement resorption. RESULTS: Kruskal-Wallis and Mann-Whitney U tests showed teeth in the negative control group to have significantly higher complete healing (98.88% +/- 2.30%) and significantly lower unfavorable healing (1.12% +/- 2.30%) than the positive control group and the experimental groups. bFGF/fibrin glue group showed higher occurrence of complete healing (39.06% +/- 41.62%) compared to the bFGF group (25.28% +/- 28.85%) and the positive control group (16.58% +/- 19.60%), although the differences were not significant. Comparing the complete and unfavorable healing, there was no significant difference in the bFGF/fibrin glue group (P = 0.47), but the differences were significant in the other groups (P < 0.05). CONCLUSION: Topical application of bFGF with fibrin glue showed an insignificantly higher occurrence of complete healing in delayed-replanted monkey teeth.     

Significance of early healing events on periodontal repair: a review.

Wound healing in an incisional wound is a highly predictable process which has been studied extensively hour-by-hour and day-by-day. Healing in a periodontal defect following gingival flap surgery is, conceptually, a more complex process as one wound margin consists of calcified tissue, including the avascular and rigid root surface. Another complicating factor in this wound healing is the transgingival position of the tooth. Experimental studies, however, have indicated that healing at a dento-gingival interface under optimal conditions occurs at the same rate as in a skin wound. Generally, periodontal healing is characterized by maturation of gingival connective tissue, limited regeneration of alveolar bone and cementum, and the formation of a long junctional epithelium. Such observations have nurtured the hypothesis that the epithelium of the surgical flap needs to be prevented from early access to the root surface during the healing period to achieve connective tissue repair of the root surface-gingival flap interface. Recent experimental findings suggest, however, that connective tissue repair to the root surface following reconstructive periodontal surgery is a function of the establishment and maintenance of a root surface-adhering fibrin clot. Since fibrin adherence to the wound margins is a natural event, it is additionally suggested that apical migration of the gingival epithelium in periodontal surgical wounds may only follow interruption of the adherence of the fibrin clot to the root surface.     

Effect of cultured autologous oral keratinocyte suspension in fibrin glue on oral wound healing in rabbits

The effect of cultured autologous oral keratinocyte suspension in fibrin glue on the healing of surgically produced oral mucosal wounds was assessed in the rabbit model. Using the light microscope and a digital image analysis system, the epithelization parameters (marginal epithelization and percentage of wound re-epithelization) were measured in haematoxylin-eosin stained sections of the wound area and compared with those of wounds treated with fibrin glue alone and untreated ones. The epithelization was significantly higher in keratinocytes plus fibrin glue-treated wounds on postoperative days 3 and 7. No significant differences were observed on postoperative day 1, when the healing process had just begun, and on postoperative day 14, when re-epithelization was completed or nearly completed in all groups. The inflammatory infiltration of the wounded mucosa was weakest in keratinocyte-treated wounds and strongest in untreated wounds. In conclusion, suspension of cultured autologous oral keratinocytes in fibrin glue significantly accelerates oral wound healing in the rabbit model and could be beneficial in the treatment of oral wounds in patients.     

Retrospective analysis of sutureless skin closure in cleft lip repair

The technique of approximating tissues resulting in minimal amount of scar usually requires skillful suturing techniques by the surgeons, especially in cleft lip repair. Increased awareness and demand for aesthetic surgical correction with quality in tissue closure has led to the invention of new materials and techniques. Amcrylate (iso amyl 2-cyanoacrylate) is retrospectively evaluated as tissue glue in cleft lip repair, and the results are compared with skin closure by 6-0 Prolene. A retrospective analysis of 60 patients with unilateral or bilateral cleft lip repair was carried out to compare the results of skin closure with Amcrylate and 6-0 Prolene. Patients were randomly divided into 2 groups, each group containing 30, and the study was designed to evaluate the quality of scars after the use of Amcrylate tissue adhesive to close the skin during cleft lip repair and its advantages over sutures (6-0 Prolene). Both groups were analyzed for the time taken for skin closure, resultant scar, parental satisfaction, and complications, and the results were found to be statistically significant for the Amcrylate group. Amcrylate, when used as tissue glue for skin closure in cleft lip repair, definitely has an edge over conventional suturing techniques.