高压氧暴露对富血小板纤维蛋白形态学作用研究
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摘要
目的探讨高压氧(hyperbaric oxygen,HBO)暴露对富血小板纤维蛋白(platelet-rich fibrin,PRF)的形态学影响。方法选30例健康志愿者抽血制备PRF,均分为3组,试验1组PRF用0.25 MPa HBO暴露1 h,试验2组用0.25 MPa HBO暴露0.5 h,对照组不用HBO暴露,分别用光学显微镜、扫描电镜(SEM)和透射电镜(TEM)观察并比较3组形态学差异。结果光学显微镜观察发现:试验1组胶原纤维染为红色,纤维排列致密,血小板多;试验2组纤维排列稍致密,血小板量较多;对照组纤维排列稀疏,血小板最少。SEM观察发现:试验1组纤维中多个血小板伸出突起,位于纤维表面或纤维之间;试验2组致密的胶原纤维交错排列,血小板伸出突起;对照组胶原纤维排列成网状,包裹多个静止的血小板。TEM观察发现:试验1组血小板内α-颗粒释放后成空泡状,在纤维蛋白或细胞之间散在颗粒样物质,白细胞中线粒体和粗面内质网增多;试验2组较多血小板α-颗粒释放,血小板内见空泡样结构;对照组血小板内α-颗粒处于激活或静止状态。结论 HBO对PRF中血小板激活有促进作用,使血小板α-颗粒释放生长因子于网状排列的胶原纤维中,发挥生物学作用。
Objective To investigate the effects of hyperbaric oxygen(HBO)exposure on the morphology of platelet-rich fibrin(PRF). Methods Thirty healthy volunteers were divided into 3 groups. The PRF of test group 1 was exposed to0.25 MPa HBO for 1 hour. The test group 2 was exposed to 0.25 MPa HBO for 0.5 h,while control group was not exposed to HBO. The morphological differences were compared respectively among the 3 groups by light microscopy,scanning electron microscopy(SEM)and transmission electron microscopy(TEM). Results Light microscope showed that the collagen fiber was stained red,the fibers were compact,and the fibers contained more platelets in test group 1. In test group 2,the fiber arrangement was relatively compact and the amount of platelets was relatively more. In the control group,collagen fibers were arranged loosely and platelet count was the least. SEM observation found that many platelets protruded in the fibers in test group 1,which were located on the surface of the fibers or between the fibers. In test group2,dense collagen fibers were interlaced and platelets protruded. The collagen fibers in the control group were arranged in a reticular form and embedded several stationary platelets. TEM showed that the α-particles in platelets of group test 1 were released and became vacuoles,which dispersed between fibrin or cells,and the mitochondria and rough endoplasmic reticulum in white blood cells increased. In the test group 2,quite a few platelet α-parti-cles were released and vacuolar structure was found in the platelets. The α-particles in platelet of the control group were in the active or stationary state. Conclusion HBO can promote the activation of platelets in PRF and make α-particles of platelets release growth factor into the reticular collagen fibers to play its biological role.
Objective To investigate the effects of hyperbaric oxygen(HBO)exposure on the morphology of platelet-rich fibrin(PRF). Methods Thirty healthy volunteers were divided into 3 groups. The PRF of test group 1 was exposed to0.25 MPa HBO for 1 hour. The test group 2 was exposed to 0.25 MPa HBO for 0.5 h,while control group was not exposed to HBO. The morphological differences were compared respectively among the 3 groups by light microscopy,scanning electron microscopy(SEM)and transmission electron microscopy(TEM). Results Light microscope showed that the collagen fiber was stained red,the fibers were compact,and the fibers contained more platelets in test group 1. In test group 2,the fiber arrangement was relatively compact and the amount of platelets was relatively more. In the control group,collagen fibers were arranged loosely and platelet count was the least. SEM observation found that many platelets protruded in the fibers in test group 1,which were located on the surface of the fibers or between the fibers. In test group2,dense collagen fibers were interlaced and platelets protruded. The collagen fibers in the control group were arranged in a reticular form and embedded several stationary platelets. TEM showed that the α-particles in platelets of group test 1 were released and became vacuoles,which dispersed between fibrin or cells,and the mitochondria and rough endoplasmic reticulum in white blood cells increased. In the test group 2,quite a few platelet α-parti-cles were released and vacuolar structure was found in the platelets. The α-particles in platelet of the control group were in the active or stationary state. Conclusion HBO can promote the activation of platelets in PRF and make α-particles of platelets release growth factor into the reticular collagen fibers to play its biological role.
引文
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[9] Chen TL,Lu HJ,Liu GQ,et al. Effect of autologous platelet-rich plasma in combination with bovine porous bone mineral and BioGuide membrane on bone regeneration in mandible bicortical bony defects[J]. J Craniofac Surg,2014,25(1):215-223.
[10] Camargo PM,Lekovic V,Weinlaender M,et al. Platelet-rich plasma and bovine porous bone mineral combined with guided tissue regeneration in the treatment of intrabony defects in humans[J]. J Periodont Res,2002,37(4):300-306.
[11] Sunitha Raja V,Munirathnam Naidu E. Platelet-rich fibrin:evaluation of a second-generation platelet concentrate[J]. Indian J Dent Res,2008,19(1):42-46.
[12] Lundquist R,Dziegiel MH,Agren MS. Bioactivity and stability of endogenous fibrogenic factors in platelet-rich fibrin[J].Wound Repair Regen,2008,16(3):356-363.
[13] Chang IC,Tsai CH,Chang YC. Platelet-rich fibrin modulates the expression of extracellular signal-regulated protein kinase and osteoprotegerin in human osteoblasts[J]. J Biomed Mater Res A,2010,95(1):327-332.
[14] Huang FM,Yang SF,Zhao JH,et al. Platelet-rich fibrin increases proliferation and differentiation of human dental pulp cells[J]. J Endod,2010,36(10):1628-1632.
[15] Handy RD,Bryson P,Moody AJ,et al. Oxidative metabolism in platelets,platelet aggregation,and hematology in patients undergoing multiple hyperbaric oxygen exposures[J]. Undersea Hyperb Med,2005,32(5):327-340.
[16] Klemetti E,Rico-Vargas S,Mojon P. Short duration hyperbaric oxygen treatment effects blood flow in rats:pilot observations[J]. Lab Anim,2005,39(1):116-121.
[17] Chen TL,Zhou YJ,Liu JC,et al. Biological effects of hyperbaric oxygen on human severe periodontitis[J]. Undersea Hyperb Med,2002,29(3):159-166.
[18]孙洁,张剑明,李彦秋.富血小板纤维蛋白超微结构的观察与探讨[J].口腔医学研究,2010,26(1):98-101.
[19] Blomb?ck B. Fibrin formation in whole blood[J]. Thromb Res,2000,99(3):307-310.
[20] Dohan DM,Choukroun J,Diss A,et al. Platelet-rich fibrin(PRF):a second generation platelet concentrate. PartⅢ:Leucocyte activation:A new feature for platelet concentrates[J].Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2006,101(3):e51-e55.
[2] Panda S,Ramamoorthi S,Jayakumar ND,et al. Platelet rich fibrin and alloplast in the treatment of intrabony defect[J]. J Pharm Bioall Sci,2014,6(2):127-131.
[3]陈铁楼,蔺世龙,刘景昌,等.高压氧对牙周炎组织前列腺素的作用及机理分析[J].中华口腔医学杂志,2002,37(3):228-230.
[4]陈铁楼,蔺世龙,许自超,等.高压氧对鼠牙龈微血管及微循环作用研究[J].微循环学杂志,1999,9(3):10-11.
[5] Jan AM,Sandor GK,Iera D,et al. Hyperbaric oxygen results in an increase in rabbit calvarial critical sized defects[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2006,101(2):144-149.
[6] Dohan DM,Choukroun J,Diss A,et al. Platelet-rich fibrin(PRF):a second generation platelet concentrate. PartⅠ:technological concepts and evolution[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2006,101(3):e37-e44.
[7]陈铁楼,刘景昌,王晓熙,等.高压氧对豚鼠实验性牙周炎作用的组织病理学研究[J].实用口腔医学杂志,2002,15(8):418-420.
[8] Dohan DM,Choukroun J,Diss A,et al. Platelet-rich fibrin(PRF):a second generation platelet concentrate. PartⅡ:platelet-related biologic features[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2006,101(3):e45-e50.
[9] Chen TL,Lu HJ,Liu GQ,et al. Effect of autologous platelet-rich plasma in combination with bovine porous bone mineral and BioGuide membrane on bone regeneration in mandible bicortical bony defects[J]. J Craniofac Surg,2014,25(1):215-223.
[10] Camargo PM,Lekovic V,Weinlaender M,et al. Platelet-rich plasma and bovine porous bone mineral combined with guided tissue regeneration in the treatment of intrabony defects in humans[J]. J Periodont Res,2002,37(4):300-306.
[11] Sunitha Raja V,Munirathnam Naidu E. Platelet-rich fibrin:evaluation of a second-generation platelet concentrate[J]. Indian J Dent Res,2008,19(1):42-46.
[12] Lundquist R,Dziegiel MH,Agren MS. Bioactivity and stability of endogenous fibrogenic factors in platelet-rich fibrin[J].Wound Repair Regen,2008,16(3):356-363.
[13] Chang IC,Tsai CH,Chang YC. Platelet-rich fibrin modulates the expression of extracellular signal-regulated protein kinase and osteoprotegerin in human osteoblasts[J]. J Biomed Mater Res A,2010,95(1):327-332.
[14] Huang FM,Yang SF,Zhao JH,et al. Platelet-rich fibrin increases proliferation and differentiation of human dental pulp cells[J]. J Endod,2010,36(10):1628-1632.
[15] Handy RD,Bryson P,Moody AJ,et al. Oxidative metabolism in platelets,platelet aggregation,and hematology in patients undergoing multiple hyperbaric oxygen exposures[J]. Undersea Hyperb Med,2005,32(5):327-340.
[16] Klemetti E,Rico-Vargas S,Mojon P. Short duration hyperbaric oxygen treatment effects blood flow in rats:pilot observations[J]. Lab Anim,2005,39(1):116-121.
[17] Chen TL,Zhou YJ,Liu JC,et al. Biological effects of hyperbaric oxygen on human severe periodontitis[J]. Undersea Hyperb Med,2002,29(3):159-166.
[18]孙洁,张剑明,李彦秋.富血小板纤维蛋白超微结构的观察与探讨[J].口腔医学研究,2010,26(1):98-101.
[19] Blomb?ck B. Fibrin formation in whole blood[J]. Thromb Res,2000,99(3):307-310.
[20] Dohan DM,Choukroun J,Diss A,et al. Platelet-rich fibrin(PRF):a second generation platelet concentrate. PartⅢ:Leucocyte activation:A new feature for platelet concentrates[J].Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2006,101(3):e51-e55.