富血小板纤维蛋白膜在急性外伤性鼓膜穿孔愈合中的应用
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摘要
目的评估富血小板纤维蛋白(platelet-rich fibrin,PRF)膜在急性外伤性鼓膜穿孔愈合中的作用。方法在64只成年雄性SD大鼠的双侧鼓膜做直径3 mm的穿孔。次日,抽取大鼠静脉血并离心,以膜的形式得到PRF,将PRF膜置于所有大鼠左侧穿孔鼓膜,为研究组;所有大鼠右侧鼓膜自然愈合,为对照组。64只大鼠随机分为8组,每组8只,分别在第3、5、7、10、14、18、22和26天肌注大剂量二甲胺和盐酸氯胺酮混合物处死。进行组织病理学检查,比较各组鼓膜穿孔最大直径、固有层水肿、新生血管的形成、成纤维细胞的生成和炎性细胞浸润等。结果研究组鼓膜穿孔最大直径小于对照组,差异有统计学意义(P<0.05)。研究组和对照组在固有层水肿、新生物血管形成、纤维化程度和炎性细胞浸润等方面的差异均无统计学意义(均P>0.05)。结论 PRF膜可加速急性外伤性鼓膜穿孔愈合。
Objective To evaluate the effect of platelet-rich fibrin(PRF)membrane in the healing process of acute tympanic membrane perforations(TMPs).Methods Acute TMPs model was established in both tympanic membranes of 64 adult male SD rats.Next day,venous blood was withdrawn from the rats,then centrifuged.PRF was obtained in membrane form.The PRF membranes were applied at the left perforated tympanic membranes of all the rats,serving as the study group.Those perforated tympanic membranes was right to spontaneously heal,as the control group.All the 64 rats were randomly divided into 8 groups(n=8 in each group)and were sacrificed on days 3,5,7,10,14,18,22 and 26,respectively,with an intramuscular injection of high-dose Xylazine and Ketamine HCl mixture.And then histopathological examination was performed.Five histopathological parameters were reviewed by 2 blinded pathologists:maximum diameter of tympanic perforations,oedema in the lamina propria,neovascularization,fibroblastic reaction,and inflammatory cell infiltration.Results The maximum diameter of tympanic perforations in the study group was smaller than that in the control group,and the difference was statistically significant(P<0.05).There were no statistically significant differences(P>0.05)between the two groups in terms of oedema in the lamina propria,neovascularization,fibroblastic reaction and inflammatory cell infiltration.Conclusion PRF fastens TMPs closure.
Objective To evaluate the effect of platelet-rich fibrin(PRF)membrane in the healing process of acute tympanic membrane perforations(TMPs).Methods Acute TMPs model was established in both tympanic membranes of 64 adult male SD rats.Next day,venous blood was withdrawn from the rats,then centrifuged.PRF was obtained in membrane form.The PRF membranes were applied at the left perforated tympanic membranes of all the rats,serving as the study group.Those perforated tympanic membranes was right to spontaneously heal,as the control group.All the 64 rats were randomly divided into 8 groups(n=8 in each group)and were sacrificed on days 3,5,7,10,14,18,22 and 26,respectively,with an intramuscular injection of high-dose Xylazine and Ketamine HCl mixture.And then histopathological examination was performed.Five histopathological parameters were reviewed by 2 blinded pathologists:maximum diameter of tympanic perforations,oedema in the lamina propria,neovascularization,fibroblastic reaction,and inflammatory cell infiltration.Results The maximum diameter of tympanic perforations in the study group was smaller than that in the control group,and the difference was statistically significant(P<0.05).There were no statistically significant differences(P>0.05)between the two groups in terms of oedema in the lamina propria,neovascularization,fibroblastic reaction and inflammatory cell infiltration.Conclusion PRF fastens TMPs closure.
引文
[1] Orji F T,Agu C C.Determinants of spontaneous healing in traumatic perforations of the tympanic membrane[J].Clin Otolaryngol,2008,33(5):420-426.
[2] Lou Z C,Lou Z H,Zhang Q P.Traumatic tympanic membrane:a study of etiology and factors affecting outcome[J].Am J Otolaryngol,2012,33(5):549-555.
[3] Hempel J M,Becker A,Muller J,et al.Traumatic tympanic membrane perforations:clinical and audiometric findings in 198 patients[J].Otol Neurotol,2012,33(8):1357-1362.
[4] Simsek G,Akin I.Early paper patching versus observation in patients with traumatic eardrum perforations:comparisons.of anatomical and functional outcomes[J].J Craniofac Surg,2014,25(6):2030-2032.
[5] Teh B M,Marano R J,Shen Y,et al.Tissue engineering of the tympanic membrane[J].Tissue Eng Part B Rev,2013,19(2):116-132.
[6] Wang A Y,Shen Y,Wang J T,et al.Animal models of chronic tympanic membrane perforation:a‘time-out’to review evidence and standardize design[J].Int J Pediatr Otorhinolaryngol,2014,78(12):2048-2055.
[7] Shen Y,Guo Y,Wilczynska M,et al Plasminogen initiates and potentiates the healing of acute and chronic tympanic membrane perforations in mice[J].J Transl Med,2014,12:5
[8] Lou Z C,Tang Y M,Chen H Y,et al.The perforation margin phenotypes and clinical outcome of traumatic tympanic membrane perforation with a Gelfoam patch:our experience from a retrospective study of seventy-four patients[J].Clin Otolaryngol,2015,40(4):389-392.
[9] Kaftan H,Herzog M,Miehe B,et al.Topical application of transforming growth factor-beta1 in acute traumatic tympanic membrane perforations:an experimental study in rats[J].Wound Repair Regen,2006,14(4):453-456.
[10] Lou Z,Yang J,Tang Y,et al.Topical application of epidermal growth factor with no scaffold material on the healing of human traumatic tympanic membrane perforations[J].Clin Otolaryngol,2016,41(6):744-749.
[11] R??sli C,von Büren T,Gassmann N B,et al.The impact of platelet-derived growth factor on closure of chronic tympanic membrane perforations:a randomized,double-blind,placebo-controlled study[J].Otol Neurotol,2011,32(8):1224-1229.
[12] Jun H J,Oh K H,Yoo J,et al.A new patch material for tympanic membrane perforation by trauma:the membrane of a hen egg shell[J].Acta Otolaryngol,2014,134(3):250-254.
[13] Barati B,Abtahi S H,Hashemi S M,et al.The effect of topical estrogen on healing of chronic tympanic membrane perforations and hearing threshold[J].J Res Med Sci,2013,18(2):99-102.
[14] Sommeling C E,Heyneman A,Hoeksema H,et al.The use of platelet-rich plasma in plastic surgery:a systematic review[J].J Plast Reconstr Aesthet Surg,2013,66(3):301-311.
[15] Kazakos K,Lyras D N,Verettas D,et al.The use of autologous PRP gel as an aid in the management of acute trauma wounds[J].Injury,2009,40(8):801-805.
[16] Saad Setta H,Elshahat A,Elsherbiny K,et al.Platelet-rich plasma versus platelet-poor plasma in the management of chronic diabetic foot ulcers:a comparative study[J].Int Wound J,2011,8(3):307-312.
[17] Knighton D R,Ciresi K,Fiegel V D,et al.Stimulation of repair in chronic,nonhealing,cutaneous ulcers using platelet-derived wound healing formula[J].Surg Gynecol Obstet,1990,170(1):56-60.
[18] Nakamura S,Ishihara M,Takikawa M,et al.Platelet-rich plasma(PRP)promotes survival of fat-grafts in rats[J].Ann Plast Surg,2010,65(1):101-106.
[19] 周阎施婧.富血小板血浆(PRP)对大鼠游离脂肪移植物存活作用的实验研究[D].武汉:华中科技大学,2012.
[20] Por Y C,Yeow V K,Louri N,et al.Platelet-rich plasma has no effect on increasing free fat graft survival in the nude mouse[J].J Plast Reconstr Aesthet Surg,2009,62(8):1030-1034.
[21] Somers T,Goovaerts G,Schelfhout L,et al.Growth factors in tympanic membrane perforations[J].Am J Otol,1998,19(4):428-434.
[22] Wang W Q,Wang Z M,Chi F L,et al.Spontaneous healing of various tympanic membrane perforations in the rat[J].Acta Otolaryngol,2004,124(10):1141-1144.
[23] Schmidt S H,Hellstrom S.Tympanic-membrane structure-new views.A comparative study[J].ORL J Otorhinolaryngol Relat Spec,1991,53(1):32-36.
[24] Lee J W,Kwon O H,Kim T K,et al.Platelet-rich plasma:quantitative assessment of growth factor levels and comparative analysis of activated and inactivated groups[J].Arch Plast Surg,2013,40(5):530-535
[25] Dohan D M,Chouknoun J,Diss A,et al.Platelet rich fibrin(PRF):a second-generation platelet concentrate.Part Ⅰ:technological concepts and evoluation[J].Oral Surg Oral Med Oral Pathol Oral Radiol Oral Endod,2006,101(3):37-44.
[26] Dohan D M,Choukroun J,Diss A,et al.Platelet-rich fibrin(PRF):a second-generation platelet concentrate.Part Ⅱ:platelet-related biologic features[J].Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2006,101(3):e45-e50.
[27] Dohan D M,Rasmusson L,Albrektsson T,et al.Classification of platelet concentrates from pure platelet-rich plasma(P-PRP)to leucocyte and platelet-rich fibrin(L-PRF)[J].Trends Biotechnol,2009,27(3):158-167.
[28] Dohan Ehrenfest D M,Bielecki T,Jimbo R,et al.Do the fibrin architecture and leukocyte content influence the growth factor release of platelet concentrates?An evidence-based answer comparing a pure platelet-rich plasma(P-PRP)gel and a leukocyte-and platelet-rich fibrin(L-PRF)[J].Curr Pharm Biotechnol,2012,13(7):1145-1152.
[29] Karatayl ?zgürsoy S,Tun?ka?k M E,Tun?ka?k F,et al.Platelet-rich plasma application for acute tympanic membrane Perforations[J].J Int Adv Otol,2017,13(2):195-199.
[30] Gür ? E,Ensari N,?ztürk M T,et al.Use of a platelet-rich fibrin membrane to repair traumatic tympanic membrane perforations:a comparative study[J].Acta Otolaryngol,2016,136(10):1017-1023.
[31] Cortese A,Pantaleo G,Borri A,et al.Platelet-rich fibrin(PRF)in implant dentistry in combination with new bone regenerative technique in elderly patients[J].Int J Surg Case Rep,2016,28:52-56.
[32] Dohan D M,Choukroun J,Diss A,et al.Platelet-rich fibrin(PRF):a second generation platelet concentrate:Part Ⅲ:leucocyte activation:a new feature for platelet concentrates?[J].Oral Surg Oral Med Oral Pathol Oral Radiol End,2006,101(3):e51-e55.
[33] Erkilet E,Koyuncu M,Atmaca S,et al.Platelet-rich plasma improves healing of tympanic membrane perforations:experimental study[J].J Laryngol Otol,2009,123(5):482-487.
[34] Goral A,Aslan C,Kucukzeybek B B,et al.Platelet-rich fibrin improves the viability of diced cartilage grafts in a rabbit model[J].Aesthet Surg,2016,36(4):153-162.
[35] Chpola S,Matta I.Cartilage-perichondrium:an ideal graft material?[J].Indian J Otolaryngol Head Neck Surg,2012,64(3):208-213.
[36] Garin P,Peerbaccus N,Mardyla F,et al.Platelet-Rich Fibrin(PRF):an autologous packing material for middle ear microsurgery[J].B-Ent 2014,10(1):27-34.
[37] Gun T,Sozen T,Boztepe O F,et al.Influence of size and site of perforation on fat graft myringoplasty[J].Auris Nasus Larynx,2014,41(6):507-512.
[38] Alvaro M L N,Ortiz N,Rodriguez L,et al.Pilot study on the efficiency of the biostimulation with autologous plasma rich in platelet growth factors in otorhinolaryngology:otologic surgery(tympanoplasty type 1)[J].ISRN Surg,2011,2011(4):451020.
[39] El-Anwar M W,El-Ahl M A,Zidan A A,et al.Topical use of autologous platelet rich plasma in myringoplasty[J].Auris Nasus Larynx,2015,42(5):365-368.
[40] Howard M L,Meyer A D,Bauer C A,et al.Middle ear tympanic membrane perforations[J].Medscape,2012.http://emedicine.medscape.com/article/858684-overview
[41] Santhi T,Rajan K V.A study of closure of tympanic membrane perforations by chemical cauterisation[J].Indian J Otolaryngol Head Neck Surg,2012,64(4):389-392.
[2] Lou Z C,Lou Z H,Zhang Q P.Traumatic tympanic membrane:a study of etiology and factors affecting outcome[J].Am J Otolaryngol,2012,33(5):549-555.
[3] Hempel J M,Becker A,Muller J,et al.Traumatic tympanic membrane perforations:clinical and audiometric findings in 198 patients[J].Otol Neurotol,2012,33(8):1357-1362.
[4] Simsek G,Akin I.Early paper patching versus observation in patients with traumatic eardrum perforations:comparisons.of anatomical and functional outcomes[J].J Craniofac Surg,2014,25(6):2030-2032.
[5] Teh B M,Marano R J,Shen Y,et al.Tissue engineering of the tympanic membrane[J].Tissue Eng Part B Rev,2013,19(2):116-132.
[6] Wang A Y,Shen Y,Wang J T,et al.Animal models of chronic tympanic membrane perforation:a‘time-out’to review evidence and standardize design[J].Int J Pediatr Otorhinolaryngol,2014,78(12):2048-2055.
[7] Shen Y,Guo Y,Wilczynska M,et al Plasminogen initiates and potentiates the healing of acute and chronic tympanic membrane perforations in mice[J].J Transl Med,2014,12:5
[8] Lou Z C,Tang Y M,Chen H Y,et al.The perforation margin phenotypes and clinical outcome of traumatic tympanic membrane perforation with a Gelfoam patch:our experience from a retrospective study of seventy-four patients[J].Clin Otolaryngol,2015,40(4):389-392.
[9] Kaftan H,Herzog M,Miehe B,et al.Topical application of transforming growth factor-beta1 in acute traumatic tympanic membrane perforations:an experimental study in rats[J].Wound Repair Regen,2006,14(4):453-456.
[10] Lou Z,Yang J,Tang Y,et al.Topical application of epidermal growth factor with no scaffold material on the healing of human traumatic tympanic membrane perforations[J].Clin Otolaryngol,2016,41(6):744-749.
[11] R??sli C,von Büren T,Gassmann N B,et al.The impact of platelet-derived growth factor on closure of chronic tympanic membrane perforations:a randomized,double-blind,placebo-controlled study[J].Otol Neurotol,2011,32(8):1224-1229.
[12] Jun H J,Oh K H,Yoo J,et al.A new patch material for tympanic membrane perforation by trauma:the membrane of a hen egg shell[J].Acta Otolaryngol,2014,134(3):250-254.
[13] Barati B,Abtahi S H,Hashemi S M,et al.The effect of topical estrogen on healing of chronic tympanic membrane perforations and hearing threshold[J].J Res Med Sci,2013,18(2):99-102.
[14] Sommeling C E,Heyneman A,Hoeksema H,et al.The use of platelet-rich plasma in plastic surgery:a systematic review[J].J Plast Reconstr Aesthet Surg,2013,66(3):301-311.
[15] Kazakos K,Lyras D N,Verettas D,et al.The use of autologous PRP gel as an aid in the management of acute trauma wounds[J].Injury,2009,40(8):801-805.
[16] Saad Setta H,Elshahat A,Elsherbiny K,et al.Platelet-rich plasma versus platelet-poor plasma in the management of chronic diabetic foot ulcers:a comparative study[J].Int Wound J,2011,8(3):307-312.
[17] Knighton D R,Ciresi K,Fiegel V D,et al.Stimulation of repair in chronic,nonhealing,cutaneous ulcers using platelet-derived wound healing formula[J].Surg Gynecol Obstet,1990,170(1):56-60.
[18] Nakamura S,Ishihara M,Takikawa M,et al.Platelet-rich plasma(PRP)promotes survival of fat-grafts in rats[J].Ann Plast Surg,2010,65(1):101-106.
[19] 周阎施婧.富血小板血浆(PRP)对大鼠游离脂肪移植物存活作用的实验研究[D].武汉:华中科技大学,2012.
[20] Por Y C,Yeow V K,Louri N,et al.Platelet-rich plasma has no effect on increasing free fat graft survival in the nude mouse[J].J Plast Reconstr Aesthet Surg,2009,62(8):1030-1034.
[21] Somers T,Goovaerts G,Schelfhout L,et al.Growth factors in tympanic membrane perforations[J].Am J Otol,1998,19(4):428-434.
[22] Wang W Q,Wang Z M,Chi F L,et al.Spontaneous healing of various tympanic membrane perforations in the rat[J].Acta Otolaryngol,2004,124(10):1141-1144.
[23] Schmidt S H,Hellstrom S.Tympanic-membrane structure-new views.A comparative study[J].ORL J Otorhinolaryngol Relat Spec,1991,53(1):32-36.
[24] Lee J W,Kwon O H,Kim T K,et al.Platelet-rich plasma:quantitative assessment of growth factor levels and comparative analysis of activated and inactivated groups[J].Arch Plast Surg,2013,40(5):530-535
[25] Dohan D M,Chouknoun J,Diss A,et al.Platelet rich fibrin(PRF):a second-generation platelet concentrate.Part Ⅰ:technological concepts and evoluation[J].Oral Surg Oral Med Oral Pathol Oral Radiol Oral Endod,2006,101(3):37-44.
[26] Dohan D M,Choukroun J,Diss A,et al.Platelet-rich fibrin(PRF):a second-generation platelet concentrate.Part Ⅱ:platelet-related biologic features[J].Oral Surg Oral Med Oral Pathol Oral Radiol Endod,2006,101(3):e45-e50.
[27] Dohan D M,Rasmusson L,Albrektsson T,et al.Classification of platelet concentrates from pure platelet-rich plasma(P-PRP)to leucocyte and platelet-rich fibrin(L-PRF)[J].Trends Biotechnol,2009,27(3):158-167.
[28] Dohan Ehrenfest D M,Bielecki T,Jimbo R,et al.Do the fibrin architecture and leukocyte content influence the growth factor release of platelet concentrates?An evidence-based answer comparing a pure platelet-rich plasma(P-PRP)gel and a leukocyte-and platelet-rich fibrin(L-PRF)[J].Curr Pharm Biotechnol,2012,13(7):1145-1152.
[29] Karatayl ?zgürsoy S,Tun?ka?k M E,Tun?ka?k F,et al.Platelet-rich plasma application for acute tympanic membrane Perforations[J].J Int Adv Otol,2017,13(2):195-199.
[30] Gür ? E,Ensari N,?ztürk M T,et al.Use of a platelet-rich fibrin membrane to repair traumatic tympanic membrane perforations:a comparative study[J].Acta Otolaryngol,2016,136(10):1017-1023.
[31] Cortese A,Pantaleo G,Borri A,et al.Platelet-rich fibrin(PRF)in implant dentistry in combination with new bone regenerative technique in elderly patients[J].Int J Surg Case Rep,2016,28:52-56.
[32] Dohan D M,Choukroun J,Diss A,et al.Platelet-rich fibrin(PRF):a second generation platelet concentrate:Part Ⅲ:leucocyte activation:a new feature for platelet concentrates?[J].Oral Surg Oral Med Oral Pathol Oral Radiol End,2006,101(3):e51-e55.
[33] Erkilet E,Koyuncu M,Atmaca S,et al.Platelet-rich plasma improves healing of tympanic membrane perforations:experimental study[J].J Laryngol Otol,2009,123(5):482-487.
[34] Goral A,Aslan C,Kucukzeybek B B,et al.Platelet-rich fibrin improves the viability of diced cartilage grafts in a rabbit model[J].Aesthet Surg,2016,36(4):153-162.
[35] Chpola S,Matta I.Cartilage-perichondrium:an ideal graft material?[J].Indian J Otolaryngol Head Neck Surg,2012,64(3):208-213.
[36] Garin P,Peerbaccus N,Mardyla F,et al.Platelet-Rich Fibrin(PRF):an autologous packing material for middle ear microsurgery[J].B-Ent 2014,10(1):27-34.
[37] Gun T,Sozen T,Boztepe O F,et al.Influence of size and site of perforation on fat graft myringoplasty[J].Auris Nasus Larynx,2014,41(6):507-512.
[38] Alvaro M L N,Ortiz N,Rodriguez L,et al.Pilot study on the efficiency of the biostimulation with autologous plasma rich in platelet growth factors in otorhinolaryngology:otologic surgery(tympanoplasty type 1)[J].ISRN Surg,2011,2011(4):451020.
[39] El-Anwar M W,El-Ahl M A,Zidan A A,et al.Topical use of autologous platelet rich plasma in myringoplasty[J].Auris Nasus Larynx,2015,42(5):365-368.
[40] Howard M L,Meyer A D,Bauer C A,et al.Middle ear tympanic membrane perforations[J].Medscape,2012.http://emedicine.medscape.com/article/858684-overview
[41] Santhi T,Rajan K V.A study of closure of tympanic membrane perforations by chemical cauterisation[J].Indian J Otolaryngol Head Neck Surg,2012,64(4):389-392.