血小板胶-胶原生物活性复合膜修复大鼠牙周骨组织缺损
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摘要
背景:近些年,组织工程学的发展为牙周组织缺损的治疗与组织再生提供了新途径。组织工程学治疗手段包括种子细胞、支架和生长因子。血小板胶因含大量血小板生长因子,胶原常被用于支架材料的制备,故血小板胶与胶原生物活性复合膜可为缺损组织提供支架及生长因子。目的:观察血小板胶-胶原生物活性复合膜在修复大鼠牙周骨组织缺损中的作用。方法:实验选取Wistar大鼠42只(由上海西普尔-必凯实验动物有限公司提供)。①将胶原裁成5mm×2mm大小,抽取6只大鼠10 mL全血后,从大鼠新鲜全血中获取富血小板血浆,按一定比例加入牛凝血酶、氯化钙和胶原制成血小板胶-胶原复合膜。测定全血及富血小板血浆中血小板计数值,ELISA法测定全血及富血小板血浆中血小板衍生生长因子AB、转化生长因子β、碱性成纤维细胞生长因子和血管内皮生长因子的水平。②将剩余36只大鼠建立下颌牙周骨缺损模型(骨缺损大小为5 mm×2 mm,磨除牙根面牙骨质)后,随机分成2组。血小板胶-胶原组将血小板胶-胶原复合膜置于骨缺损处,而对照组不放置任何材料。2,4和8周后获取2组大鼠牙周组织标本进行苏木精-伊红染色,测定新生牙周骨率、新生牙骨质、新生牙槽骨、新生牙周膜组织高度。免疫组织化学染色观察骨形成蛋白2的表达。结果与结论:①富血小板血浆中血小板计数均值为全血的4.78倍,即全血制备成富血小板血浆后血小板数量显著上升(P<0.05)。富血小板血浆中血小板衍生生长因子AB、转化生长因子β、碱性成纤维细胞生长因子和血管内皮生长因子的水平分别是全血中的3.10倍、3.45倍、7.17倍、5.45倍(P<0.05); ②苏木精-伊红染色观察结果:两组新生骨率、新生牙骨质、新生牙槽骨、新生牙周膜组织高度在2周时差异无显著性意义(P> 0.05),第4和第8周时,血小板胶-胶原组上述指标均显著高于对照组(P <0.05);③免疫组化结果显示,在2周时,血小板胶-胶原组骨形态发生蛋白2开始表达,4周时骨形态发生蛋白2表达最强(P <0.05),而8周时阳性表达减弱(P>0.05);④结果提示,血小板胶-胶原生物活性复合膜可明显促进新生牙组织再生,并伴随着骨形态发生蛋白2的表达,减小牙周组织缺损后的修复时间。
BACKGROUND: In recent years, the development of tissue engineering has provided a new approach for the treatment of periodontal bone defect. Tissue engineering therapy includes seed cells, scaffolds and growth factors. Platelet gel contains a large number of platelet growth factors, and collagen is often used for the preparation of scaffold materials. Therefore, the platelet gel and collagen biologically active composite membrane can provide scaffolds and growth factors for the defect bone. OBJECTIVE: To investigate the effect of autologous platelet gel-collagen biologically active composite membrane on the repair of periodontal bone defect in rats. METHODS: Forty-two Wistar rats(Shanghai Xipuer-Bikai Experimental Animal Co., Ltd., China) were selected.(1) Collagen was cut into 5 mm×2 mm size, and 10 mL of whole blood was extracted from 6 rats to obtain platelet-rich plasma. Autologous platelet gel-collagen composite membrane was prepared by adding bovine thrombin, calcium chloride and collagen in a certain proportion. Platelets in whole blood and in platelet-rich plasma were detected. The levels of platelet derived growth factor AB, transforming growth factor-β, basic fibroblast growth factor and vascular endothelial growth factor in whole blood and platelet-rich plasma were detected by ELISA.(2) The models of mandibular periosteal defect were established in 36 rats(the size of the bone defect was 5 mm×2 mm, and the root surface cementum was removed), and randomly divided into two groups. Autologous platelet gel-collagen group placed the autologous platelet gel-collagen composite membrane in the bone defect, and the control group did not place any materials. The hematoxylin-eosin staining of periodontal tissues of rats in each group was analyzed at 2, 4 and 8 weeks after surgery. Rate of new born, new centumum formation, new alveolar bone formation, and new periodontal ligament tissue formation height were measured. The expression of bone morphogenetic protein-2 was detected by immunohistochemical staining. RESULTS AND CONCLUSION:(1) The mean platelet count in platelet-rich plasma was 4.78 times as high as the whole blood, indicating that the number of platelets increased significantly after prepared into platelet-rich plasma(P < 0.05). The levels of platelet derived growth factor AB, transforming growth factor-β, basic fibroblast growth factor and vascular endothelial growth factor in platelet-rich plasma were 3.10, 3.45, 7.17 and 5.45 times of the whole blood, respectively(P < 0.05).(2) The results of hematoxylin-eosin staining observed that the rate of new born, new centumum formation, new alveolar bone formation, and new periodontal ligament tissue formation height at 2 weeks in the autologous platelet gel-collagen group showed no significant difference from the control group(P > 0.05). At 4 and 8 weeks, all above indexes in the autologous platelet gel-collagen group were significantly higher than those in the control group(P < 0.05).(3) Results of immunohistochemical staining revealed that at 2 weeks, bone morphogenetic protein-2 in the autologous platelet gel-collagen group began to express, and the expression of bone morphogenetic protein-2 was highest at 4 weeks(P < 0.05), and the positive expression was weakened at 8 weeks(P > 0.05).(4) Our results clarify that autologous platelet gel-collagen bioactive composite membrane can significantly promote the regeneration of new tooth, which is associated with the expression of bone morphogenetic protein-2, and reduce the repair time after periodontal tissue defect.
BACKGROUND: In recent years, the development of tissue engineering has provided a new approach for the treatment of periodontal bone defect. Tissue engineering therapy includes seed cells, scaffolds and growth factors. Platelet gel contains a large number of platelet growth factors, and collagen is often used for the preparation of scaffold materials. Therefore, the platelet gel and collagen biologically active composite membrane can provide scaffolds and growth factors for the defect bone. OBJECTIVE: To investigate the effect of autologous platelet gel-collagen biologically active composite membrane on the repair of periodontal bone defect in rats. METHODS: Forty-two Wistar rats(Shanghai Xipuer-Bikai Experimental Animal Co., Ltd., China) were selected.(1) Collagen was cut into 5 mm×2 mm size, and 10 mL of whole blood was extracted from 6 rats to obtain platelet-rich plasma. Autologous platelet gel-collagen composite membrane was prepared by adding bovine thrombin, calcium chloride and collagen in a certain proportion. Platelets in whole blood and in platelet-rich plasma were detected. The levels of platelet derived growth factor AB, transforming growth factor-β, basic fibroblast growth factor and vascular endothelial growth factor in whole blood and platelet-rich plasma were detected by ELISA.(2) The models of mandibular periosteal defect were established in 36 rats(the size of the bone defect was 5 mm×2 mm, and the root surface cementum was removed), and randomly divided into two groups. Autologous platelet gel-collagen group placed the autologous platelet gel-collagen composite membrane in the bone defect, and the control group did not place any materials. The hematoxylin-eosin staining of periodontal tissues of rats in each group was analyzed at 2, 4 and 8 weeks after surgery. Rate of new born, new centumum formation, new alveolar bone formation, and new periodontal ligament tissue formation height were measured. The expression of bone morphogenetic protein-2 was detected by immunohistochemical staining. RESULTS AND CONCLUSION:(1) The mean platelet count in platelet-rich plasma was 4.78 times as high as the whole blood, indicating that the number of platelets increased significantly after prepared into platelet-rich plasma(P < 0.05). The levels of platelet derived growth factor AB, transforming growth factor-β, basic fibroblast growth factor and vascular endothelial growth factor in platelet-rich plasma were 3.10, 3.45, 7.17 and 5.45 times of the whole blood, respectively(P < 0.05).(2) The results of hematoxylin-eosin staining observed that the rate of new born, new centumum formation, new alveolar bone formation, and new periodontal ligament tissue formation height at 2 weeks in the autologous platelet gel-collagen group showed no significant difference from the control group(P > 0.05). At 4 and 8 weeks, all above indexes in the autologous platelet gel-collagen group were significantly higher than those in the control group(P < 0.05).(3) Results of immunohistochemical staining revealed that at 2 weeks, bone morphogenetic protein-2 in the autologous platelet gel-collagen group began to express, and the expression of bone morphogenetic protein-2 was highest at 4 weeks(P < 0.05), and the positive expression was weakened at 8 weeks(P > 0.05).(4) Our results clarify that autologous platelet gel-collagen bioactive composite membrane can significantly promote the regeneration of new tooth, which is associated with the expression of bone morphogenetic protein-2, and reduce the repair time after periodontal tissue defect.
引文
[1]Moradian H,Rafiee A,Ayatollahi M.Design and fabrication of a novel transplant combined with human bone marrow mesenchymal stem cells and platelet-rich fibrin:new horizons for periodontal tissue regeneration after dental trauma.Iran J Pharm Res.2017;16(4):1370-1378.
[2]Jimbo R,Singer J,Tovar N,et al.Regeneration of the cementum and periodontal ligament using local BDNF delivery in class II furcation defects.J Biomed Mater Res B Appl Biomater.2018;106(4):1611-1617.
[3]Ogawa K,Miyaji H,Kato A,et al.Periodontal tissue engineering by nano beta-tricalcium phosphate scaffold and fibroblast growth factor-2 in one-wall infrabony defects of dogs.J Periodontal Res.2016;51(6):758-767.
[4]Heng NH,Zahlten J,Cordes V,et al.Effects of enamel matrix derivative and transforming growth factor-β1 on connective tissue growth factor in human periodontal ligament fibroblasts.J Periodontol,.2015;86(4):569-577.
[5]Rakhmatia YD,Ayukawa Y,Furuhashi A,et al.Current barrier membranes:titanium mesh and other membranes for guided bone regeneration in dental applications.J Prosthodont Res.2013;57(1):3-14.
[6]陈凤超,陈民才,颜彤彤,等.同种异体富血小板血浆对创面胶原合成的作用及其机制研究[J].中华外科杂志,2017,55(4):303-307.
[7]李晋,王秉新.Bio-Oss结合Bio-Gide修复牙槽临界骨缺损的效果[J].中国老年学杂志,2013,33(15):3666-3667.
[8]Duruel T,?akmak AS,Akman A,et al.Sequential IGF-1 and BMP-6releasing chitosan/alginate/PLGA hybrid scaffolds for periodontal regeneration.Int J Biol Macromol.2017;104(Pt A):232-241.
[9]Dhurat R,Sukesh M.Principles and methods of preparation of platelet-rich plasma:a review and author's perspective.J Cutan Aesthet Surg.2014;7(4):189-197.
[10]Takeuchi T,Bizenjima T,Ishii Y,et al.Enhanced healing of surgical periodontal defects in rats following application of a self-assembling peptide nanofibre hydrogel.J Clin Periodontol.2016;43(3):279-288.
[11]李睿,王春兰,范月静,等.纳米胶原基骨修复牙周病及根尖周病骨缺损[J].中国组织工程研究,2012,16(34):6317-6320.
[12]詹雅琳,胡文杰,徐涛,等.罹患重度牙周炎磨牙拔除后应用去蛋白牛骨基质与可吸收胶原膜进行位点保存的组织学研究[J].北京大学学报(医学版),2017,49(1):169-175.
[13]Zhou T,Liu X,Sui B,et al.Development of fish collagen/bioactive glass/chitosan composite nanofibers as a GTR/GBR membrane for inducing periodontal tissue regeneration.Biomed Mater.2017;12(5):055004.
[14]Hudson DM,Garibov M,Dixon DR,et al.Distinct post-translational features of type I collagen are conserved in mouse and humanperiodontal ligament.J Periodontal Res.2017;52(6):1042-1049.
[15]Sartori M,Giavaresi G,Parrilli A,et al.Collagen type I coating stimulates bone regeneration and osteointegration of titanium implants in the osteopenic rat.Int Orthop.2015;39(10):2041-2052.
[16]Capella-Monsonís H,Coentro JQ,Graceffa V,et al.An experimental toolbox for characterization of mammalian collagen type I in biological specimens.Nat Protoc.2018;13(3):507-529.
[17]Jo JY,Jeong SI,Shin YM,et al.Sequential delivery of BMP-2 and BMP-7for bone regeneration using a heparinized collagen membrane.Int J Oral Maxillofac Surg.2015;44(7):921-928.
[18]De Pascale MR,Sommese L,Casamassimi A,et al.Platelet derivatives in regenerative medicine:an update.Transfus Med Rev.2015;29(1):52-61.
[19]楚佳奇,孙杰聪,李鹏,等.富血小板血浆联合人脐带间充质干细胞移植对大鼠骨质疏松骨折愈合的影响[J].中国骨质疏松杂志,2016,22(3):283-287.
[20]李君,梁刘凤,钟雯怡,等.羊富血小板胶细胞基质在2种牙周缺损模型中的修复效果[J].口腔材料器械杂志,2016,25(4):204-208.
[21]Yu T,Pan H,Hu Y,et al.Autologous platelet-rich plasma induces bone formation of tissue-engineered bone with bone marrow mesenchymal stem cells on beta-tricalcium phosphate ceramics.J Orthop Surg Res.2017;12(1):178.
[22]Shafieian R,Matin MM,Rahpeyma A,et al.Effects of human adipose-derived stem cells and platelet-rich plasma on healing response of canine alveolar surgical bone defects.Arch Bone Jt Surg.2017;5(6):406-418.
[23]De Pascale MR,Sommese L,Casamassimi A,et al.Platelet derivatives in regenerative medicine:an update.Transfus Med Rev.2015;29(1):52-61.
[24]Willemsen JC,van der Lei B,Vermeulen KM,et al.The effects of platelet-rich plasma on recovery time and aesthetic outcome in facial rejuvenation:preliminary retrospective observations.Aesthetic Plast Surg.2014;38(5):1057-1063.
[25]Piccin A,Rebulla P,Pupella S,et al.Impressive tissue regeneration of severe oral mucositis post stem cell transplantation using cord blood platelet gel.Transfusion.2017;57(9):2220-2224.
[26]Mori M,Rossi S,Bonferoni MC,et al.Calcium alginate particles for the combined delivery of platelet lysate and vancomycin hydrochloride in chronic skin ulcers.Int J Pharm.2014;30,461(1-2):505-513.
[27]王淑君.血小板胶促进创面愈合的研究进展[J].医学研究生学报,2017,30(3):333-336.
[28]Fioravanti C,Frustaci I,Armellin E,et al.Autologous blood preparations rich in platelets,fibrin and growth factors.Oral Implantol(Rome).2016;8(4):96-113.
[29]Yamano S,Haku K,Yamanaka T,et al.The effect of a bioactive collagen membrane releasing PDGF or GDF-5 on bone regeneration.Biomaterials.2014;35(8):2446-2453.
[30]Stampanoni Bassi M,Iezzi E,Marfia GA,et al.Platelet-derived growth factor predicts prolonged relapse-free period in multiple sclerosis.JNeuroinflammation.2018;15(1):108.
[31]Torres K,Pietrzyk?,Plewa Z,et al.TGF-βand inflammatory blood markers in prediction of intraperitoneal adhesions.Adv Med Sci.2017;63(2):220-223.
[32]顾涛,张彦秋,刘立杰,等.非小细胞肺癌患者化疗前后血清VEGF/血小板数和TGF-β1检测的临床意义[J].中国实验诊断学,2017,21(4):585-588.
[33]Oryan A,Moshiri A,Meimandi-Parizi A,et al.Role of xenogenous bovine platelet gel embedded within collagen implant on tendon healing:an in vitro and in vivo study.Exp Biol Med(Maywood).2015;240(2):194-210.
[34]Kobayashi E,Flückiger L,Fujioka-Kobayashi M,et al.Comparative release of growth factors from PRP,PRF and advanced-PRF.Clin Oral Investig.2016;20(9):2353-2360.
[35]林静,陈超,林敏魁,等.牙周组织工程中动物模型的研究进展[J].全科口腔医学电子杂志,2016,3(20):10-12.
[36]Kaku M,Kitami M,Rosales Rocabado JM,et al.Recruitment of bone marrow-derived cells to the periodontal ligament via the stromal cell-derived factor-1/C-X-C chemokine receptor type 4 axis.JPeriodontal Res.2017;52(4):686-694.
[37]Scioli MG,Bielli A,Gentile P,et al.Combined treatment with platelet-rich plasma and insulin favours chondrogenic and osteogenic differentiation of human adipose-derived stem cells in three-dimensional collagen scaffolds.J Tissue Eng Regen Med.2017;11(8):2398-2410.
[38]许繁,杨德圣.Bio-Gide胶原膜对兔骨髓间充质干细胞增殖和成骨分化的影响[J].中国组织工程研究,2014,18(30):4797-4803.
[39]Holmes HL,Wilson B,Goerger JP,et al.Facilitated recruitment of mesenchymal stromal cells by bone marrow concentrate and platelet rich plasma.PLoS One.2018;13(3):e0194567.
[40]Hirakawa Y,Manaka T,Orita K,et al.The accelerated effect of recombinant human bone morphogenetic protein 2 delivered byβ-tricalcium phosphate on tendon-to-bone repair process in rabbit models.J Shoulder Elbow Surg.2018;27(5):894-902.
[2]Jimbo R,Singer J,Tovar N,et al.Regeneration of the cementum and periodontal ligament using local BDNF delivery in class II furcation defects.J Biomed Mater Res B Appl Biomater.2018;106(4):1611-1617.
[3]Ogawa K,Miyaji H,Kato A,et al.Periodontal tissue engineering by nano beta-tricalcium phosphate scaffold and fibroblast growth factor-2 in one-wall infrabony defects of dogs.J Periodontal Res.2016;51(6):758-767.
[4]Heng NH,Zahlten J,Cordes V,et al.Effects of enamel matrix derivative and transforming growth factor-β1 on connective tissue growth factor in human periodontal ligament fibroblasts.J Periodontol,.2015;86(4):569-577.
[5]Rakhmatia YD,Ayukawa Y,Furuhashi A,et al.Current barrier membranes:titanium mesh and other membranes for guided bone regeneration in dental applications.J Prosthodont Res.2013;57(1):3-14.
[6]陈凤超,陈民才,颜彤彤,等.同种异体富血小板血浆对创面胶原合成的作用及其机制研究[J].中华外科杂志,2017,55(4):303-307.
[7]李晋,王秉新.Bio-Oss结合Bio-Gide修复牙槽临界骨缺损的效果[J].中国老年学杂志,2013,33(15):3666-3667.
[8]Duruel T,?akmak AS,Akman A,et al.Sequential IGF-1 and BMP-6releasing chitosan/alginate/PLGA hybrid scaffolds for periodontal regeneration.Int J Biol Macromol.2017;104(Pt A):232-241.
[9]Dhurat R,Sukesh M.Principles and methods of preparation of platelet-rich plasma:a review and author's perspective.J Cutan Aesthet Surg.2014;7(4):189-197.
[10]Takeuchi T,Bizenjima T,Ishii Y,et al.Enhanced healing of surgical periodontal defects in rats following application of a self-assembling peptide nanofibre hydrogel.J Clin Periodontol.2016;43(3):279-288.
[11]李睿,王春兰,范月静,等.纳米胶原基骨修复牙周病及根尖周病骨缺损[J].中国组织工程研究,2012,16(34):6317-6320.
[12]詹雅琳,胡文杰,徐涛,等.罹患重度牙周炎磨牙拔除后应用去蛋白牛骨基质与可吸收胶原膜进行位点保存的组织学研究[J].北京大学学报(医学版),2017,49(1):169-175.
[13]Zhou T,Liu X,Sui B,et al.Development of fish collagen/bioactive glass/chitosan composite nanofibers as a GTR/GBR membrane for inducing periodontal tissue regeneration.Biomed Mater.2017;12(5):055004.
[14]Hudson DM,Garibov M,Dixon DR,et al.Distinct post-translational features of type I collagen are conserved in mouse and humanperiodontal ligament.J Periodontal Res.2017;52(6):1042-1049.
[15]Sartori M,Giavaresi G,Parrilli A,et al.Collagen type I coating stimulates bone regeneration and osteointegration of titanium implants in the osteopenic rat.Int Orthop.2015;39(10):2041-2052.
[16]Capella-Monsonís H,Coentro JQ,Graceffa V,et al.An experimental toolbox for characterization of mammalian collagen type I in biological specimens.Nat Protoc.2018;13(3):507-529.
[17]Jo JY,Jeong SI,Shin YM,et al.Sequential delivery of BMP-2 and BMP-7for bone regeneration using a heparinized collagen membrane.Int J Oral Maxillofac Surg.2015;44(7):921-928.
[18]De Pascale MR,Sommese L,Casamassimi A,et al.Platelet derivatives in regenerative medicine:an update.Transfus Med Rev.2015;29(1):52-61.
[19]楚佳奇,孙杰聪,李鹏,等.富血小板血浆联合人脐带间充质干细胞移植对大鼠骨质疏松骨折愈合的影响[J].中国骨质疏松杂志,2016,22(3):283-287.
[20]李君,梁刘凤,钟雯怡,等.羊富血小板胶细胞基质在2种牙周缺损模型中的修复效果[J].口腔材料器械杂志,2016,25(4):204-208.
[21]Yu T,Pan H,Hu Y,et al.Autologous platelet-rich plasma induces bone formation of tissue-engineered bone with bone marrow mesenchymal stem cells on beta-tricalcium phosphate ceramics.J Orthop Surg Res.2017;12(1):178.
[22]Shafieian R,Matin MM,Rahpeyma A,et al.Effects of human adipose-derived stem cells and platelet-rich plasma on healing response of canine alveolar surgical bone defects.Arch Bone Jt Surg.2017;5(6):406-418.
[23]De Pascale MR,Sommese L,Casamassimi A,et al.Platelet derivatives in regenerative medicine:an update.Transfus Med Rev.2015;29(1):52-61.
[24]Willemsen JC,van der Lei B,Vermeulen KM,et al.The effects of platelet-rich plasma on recovery time and aesthetic outcome in facial rejuvenation:preliminary retrospective observations.Aesthetic Plast Surg.2014;38(5):1057-1063.
[25]Piccin A,Rebulla P,Pupella S,et al.Impressive tissue regeneration of severe oral mucositis post stem cell transplantation using cord blood platelet gel.Transfusion.2017;57(9):2220-2224.
[26]Mori M,Rossi S,Bonferoni MC,et al.Calcium alginate particles for the combined delivery of platelet lysate and vancomycin hydrochloride in chronic skin ulcers.Int J Pharm.2014;30,461(1-2):505-513.
[27]王淑君.血小板胶促进创面愈合的研究进展[J].医学研究生学报,2017,30(3):333-336.
[28]Fioravanti C,Frustaci I,Armellin E,et al.Autologous blood preparations rich in platelets,fibrin and growth factors.Oral Implantol(Rome).2016;8(4):96-113.
[29]Yamano S,Haku K,Yamanaka T,et al.The effect of a bioactive collagen membrane releasing PDGF or GDF-5 on bone regeneration.Biomaterials.2014;35(8):2446-2453.
[30]Stampanoni Bassi M,Iezzi E,Marfia GA,et al.Platelet-derived growth factor predicts prolonged relapse-free period in multiple sclerosis.JNeuroinflammation.2018;15(1):108.
[31]Torres K,Pietrzyk?,Plewa Z,et al.TGF-βand inflammatory blood markers in prediction of intraperitoneal adhesions.Adv Med Sci.2017;63(2):220-223.
[32]顾涛,张彦秋,刘立杰,等.非小细胞肺癌患者化疗前后血清VEGF/血小板数和TGF-β1检测的临床意义[J].中国实验诊断学,2017,21(4):585-588.
[33]Oryan A,Moshiri A,Meimandi-Parizi A,et al.Role of xenogenous bovine platelet gel embedded within collagen implant on tendon healing:an in vitro and in vivo study.Exp Biol Med(Maywood).2015;240(2):194-210.
[34]Kobayashi E,Flückiger L,Fujioka-Kobayashi M,et al.Comparative release of growth factors from PRP,PRF and advanced-PRF.Clin Oral Investig.2016;20(9):2353-2360.
[35]林静,陈超,林敏魁,等.牙周组织工程中动物模型的研究进展[J].全科口腔医学电子杂志,2016,3(20):10-12.
[36]Kaku M,Kitami M,Rosales Rocabado JM,et al.Recruitment of bone marrow-derived cells to the periodontal ligament via the stromal cell-derived factor-1/C-X-C chemokine receptor type 4 axis.JPeriodontal Res.2017;52(4):686-694.
[37]Scioli MG,Bielli A,Gentile P,et al.Combined treatment with platelet-rich plasma and insulin favours chondrogenic and osteogenic differentiation of human adipose-derived stem cells in three-dimensional collagen scaffolds.J Tissue Eng Regen Med.2017;11(8):2398-2410.
[38]许繁,杨德圣.Bio-Gide胶原膜对兔骨髓间充质干细胞增殖和成骨分化的影响[J].中国组织工程研究,2014,18(30):4797-4803.
[39]Holmes HL,Wilson B,Goerger JP,et al.Facilitated recruitment of mesenchymal stromal cells by bone marrow concentrate and platelet rich plasma.PLoS One.2018;13(3):e0194567.
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