Choukroun富血小板纤维蛋白在拔牙位点保存中应用的实验研究
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摘要
牙齿拔除后,由于生理性刺激的丧失,牙槽嵴骨质会发生吸收,这种进行性、不可逆性的吸收常常会造成牙槽嵴的低平及其原有形态的改变,从而严重影响了义齿的修复,更不利于种植体的植入,也会进一步影响种植修复后的美观效果。目前,国内外专家一直致力于寻找一种有效减少拔牙后牙槽嵴吸收的方法。新一代的血小板浓缩制品即Choukroun富血小板纤维蛋白(Choukroun’s platelet-rich fibrin,Choukroun's PRF)制备方法简便,取材快捷,纤维蛋白的结构和其与细胞因子之间的协同作用优于以往的血目前小板浓缩物。,PRF已应用于多种领域。
本实验通过在兔的上、下颌切牙拔牙窝内分别植入PRF、羟基磷灰石生物陶瓷(骨粉)以及空白对照,对比观察PRF对拔牙窝牙槽嵴修复的能力,并以此为理论基础,指导其应用于拔牙位点保存的临床实践。
通过将PRF独立充填于大耳白兔拔牙窝内,观察其对拔牙窝修复的作用发现:在牙槽嵴愈合修复的早期过程中,PRF在拔牙窝骨缺损区形成纤维网络支架,促进细胞的迁移、增殖和分化,显著改善拔牙窝新骨的形成与改建,同时有效的减轻炎症水肿反应,发挥炎症调节的作用。
Background and Objective
The normal metabolism of the alveolar bone depends on the functional stimulation of teeth.If the teeth are pulled out for some reasons, the stimulus way changes and the alveolar bone will suffer persistent, progressive and non-reversible resorption,which makes the height reduced and the width narrowed down. The original form of alveolar ridge varies,which not only seriously affects the function of dentures , but also does harm to the implantation and aesthetics.
The restoration of teeth extraction is complicated.The mechanism of which is still uncertain. Many experts have been in search of how to prevent the alveolar bone from being absorbed and accelerate the restoration,then people can preserve the original shape of extraction. Scholars tried to maintain the height and width of the alveolar ridge by implanting the biological materials with a function of bone guiding and the complex of growth factors which plays a critical role in bone induction into the extraction socket.This method has become a tendency due to its stable effect and having no trauma. Choukroun's platelet-rich fibrin (PRF) is a new generation of platelet concentrates which has been extracted by Choukroun and other French scientists in 2000. There are several advantages as follows:Simple preparation that can be manipulated before or during the surgery without any additives like anti-coagulate agents or blood preparation won't induce any cross contamination or immunologic rejection,and the effective process of simulating the physiological state of the fibrin clot formation makes it similar to the normal fibrin in the human blood clot. All kinds of cell growth factors in it not only increase the concentration of cytokines,but also promote the synergistic action among them. In recent years, both basic and clinical researches suggested that the PRF has a good effect on tissue healing, inflammatory regulating and anti-infection.
The polymerization of PRF is slow and nature in the low-speed centrifu- gation state,which makes the fibrin a 3-dimensional organization and a more porous gel with larger pores and elasticity.As a substrate,PRF provides a favorable place for cells adhesion,migration and differentiation,those cytokines combine with the substrate in chemical bonds . The mechanisms of the tissue healing can be summarized as follows:1) to guide revascularization in the damaged tissue; 2) to guide epithelial tissue to close injuries;3) immunologic regulation; 4) to condut circulating blood stem cells to migrate,proliferate and differentiate.
In this study, according to the resorption and atrophy of the alveolar ridge caused by teeth extraction and the effect of PRF in tissue healing, we compared the restoration of the remaining alveolar bone by implanting the PRF, hydro- xyapatite in the sockets and the blank control after teeth extraction by means of general observation, histological observation and radiation measurement from animal experimental studies and hoped to provide some guiding significance of PRF in the clinical application.
Methods
After 2 weeks of feeding,we ground the right upper and lower incisors of the rabbits to 1 / 2 of their labial length for four times with the interval of 4 days.In the process ,the marrow were protected from being wore to avoid the infection. Then,we got the autologous blood of the animals through the ear marginal artery instantly and made it centrifuged to prepare the PRF membrane.After general anesthesia by SumianxinⅡand local anesthesia around the mucosa,we pulled out the right upper and lower incisors. We filled PRF membrane into the experimental sockets and hydroxyapatite into the positive control one ,both of which are flush with the alveolar crest level. Then we sorted out the sockets of the blank group and made it full of blood. Gingiva were tightly sutured to closed the teeth sockets in three groups. Respectively after 1 week, 2 weeks, 4 weeks, 8 weeks and 12 weeks animals were killed ,then we produced histological specimens for general observation, X-ray and HE staining microscopic observation.We calculated the ratio of the length between the extraction and non-extraction side on the X-ray films by Digora For Windows2.5 software,and disposed the data with statistical software SPSS 13.0.
Results
Generally we observed that there were no obvious infection and material-off phenomenon in the dental extraction.It belongs to primary bone healing.There is no difference in the surface healing among the the experimental group (PRF), the positive control group (bone meal group) and the negative control group (blank group).1-3 days after teeth extraction, the mucosa of the positive control groups and negative control groups appeared slight red,while the experimental group 's was normal. The surface of extraction wounds were completely covered by epithelial tissue after five days .
The relative length of the remaining alveolar ridge observed by X-ray resulted as follows: the experimental group (PRF)> the positive control group (bone meal group) >the negative control group (control group), the difference was statistically significant ( P <0.05) after 1 week, 2 weeks and 4 weeks. After 8 weeks, 12 weeks, the experimental group (PRF) and the positive control group (bone meal group) are greater than the negative control group (control group)(P<0.05) .But there was no significant difference between the experi- mental group (PRF) and the positive control group (bone meal group)( P >0.05).
The quantity and quality of new bone in the sockets of the experimental group (PRF ) were better than the positive control group (bone meal) and negative control group in the histological observation of 1 week, 2 weeks and 4 weeks . There was a large number of new bone along the nest wall which grew to the center of the sockets in the experimental group in 1 week and the experi- mental group had more lacunae and active osteoblasts than the positive control and the negative control group.There was still new bone formed in the positive control group (bone meal group) after 8 and 12 weeks,however,of which the experimental group (PRF group) and positive control group (bone meal group) had been completed remodelled, there was no significant difference on the extent of calcification in the bone lamellar structure between the two groups.
Conclusion
In the experimental group,we compared the restoration of the remaining alveolar bone by implanting the PRF ,hydroxyapatite in the sockets and the blank control after teeth extraction.All in all,the conclusions were as follows:
1、PRF and hydroxyapatite bioceramics (bone meal) can all preserve the height of the alveolar ridge after teeth extraction while PRF is more evident at an early stage.
2、PRF can restore the alveolar ridge as a independent transplant material.
34、PRF can remedy the inflammatory symptoms which can reduce post-traumatic inflammation and accelerate the healing process of wound.
本实验通过在兔的上、下颌切牙拔牙窝内分别植入PRF、羟基磷灰石生物陶瓷(骨粉)以及空白对照,对比观察PRF对拔牙窝牙槽嵴修复的能力,并以此为理论基础,指导其应用于拔牙位点保存的临床实践。
通过将PRF独立充填于大耳白兔拔牙窝内,观察其对拔牙窝修复的作用发现:在牙槽嵴愈合修复的早期过程中,PRF在拔牙窝骨缺损区形成纤维网络支架,促进细胞的迁移、增殖和分化,显著改善拔牙窝新骨的形成与改建,同时有效的减轻炎症水肿反应,发挥炎症调节的作用。
Background and Objective
The normal metabolism of the alveolar bone depends on the functional stimulation of teeth.If the teeth are pulled out for some reasons, the stimulus way changes and the alveolar bone will suffer persistent, progressive and non-reversible resorption,which makes the height reduced and the width narrowed down. The original form of alveolar ridge varies,which not only seriously affects the function of dentures , but also does harm to the implantation and aesthetics.
The restoration of teeth extraction is complicated.The mechanism of which is still uncertain. Many experts have been in search of how to prevent the alveolar bone from being absorbed and accelerate the restoration,then people can preserve the original shape of extraction. Scholars tried to maintain the height and width of the alveolar ridge by implanting the biological materials with a function of bone guiding and the complex of growth factors which plays a critical role in bone induction into the extraction socket.This method has become a tendency due to its stable effect and having no trauma. Choukroun's platelet-rich fibrin (PRF) is a new generation of platelet concentrates which has been extracted by Choukroun and other French scientists in 2000. There are several advantages as follows:Simple preparation that can be manipulated before or during the surgery without any additives like anti-coagulate agents or blood preparation won't induce any cross contamination or immunologic rejection,and the effective process of simulating the physiological state of the fibrin clot formation makes it similar to the normal fibrin in the human blood clot. All kinds of cell growth factors in it not only increase the concentration of cytokines,but also promote the synergistic action among them. In recent years, both basic and clinical researches suggested that the PRF has a good effect on tissue healing, inflammatory regulating and anti-infection.
The polymerization of PRF is slow and nature in the low-speed centrifu- gation state,which makes the fibrin a 3-dimensional organization and a more porous gel with larger pores and elasticity.As a substrate,PRF provides a favorable place for cells adhesion,migration and differentiation,those cytokines combine with the substrate in chemical bonds . The mechanisms of the tissue healing can be summarized as follows:1) to guide revascularization in the damaged tissue; 2) to guide epithelial tissue to close injuries;3) immunologic regulation; 4) to condut circulating blood stem cells to migrate,proliferate and differentiate.
In this study, according to the resorption and atrophy of the alveolar ridge caused by teeth extraction and the effect of PRF in tissue healing, we compared the restoration of the remaining alveolar bone by implanting the PRF, hydro- xyapatite in the sockets and the blank control after teeth extraction by means of general observation, histological observation and radiation measurement from animal experimental studies and hoped to provide some guiding significance of PRF in the clinical application.
Methods
After 2 weeks of feeding,we ground the right upper and lower incisors of the rabbits to 1 / 2 of their labial length for four times with the interval of 4 days.In the process ,the marrow were protected from being wore to avoid the infection. Then,we got the autologous blood of the animals through the ear marginal artery instantly and made it centrifuged to prepare the PRF membrane.After general anesthesia by SumianxinⅡand local anesthesia around the mucosa,we pulled out the right upper and lower incisors. We filled PRF membrane into the experimental sockets and hydroxyapatite into the positive control one ,both of which are flush with the alveolar crest level. Then we sorted out the sockets of the blank group and made it full of blood. Gingiva were tightly sutured to closed the teeth sockets in three groups. Respectively after 1 week, 2 weeks, 4 weeks, 8 weeks and 12 weeks animals were killed ,then we produced histological specimens for general observation, X-ray and HE staining microscopic observation.We calculated the ratio of the length between the extraction and non-extraction side on the X-ray films by Digora For Windows2.5 software,and disposed the data with statistical software SPSS 13.0.
Results
Generally we observed that there were no obvious infection and material-off phenomenon in the dental extraction.It belongs to primary bone healing.There is no difference in the surface healing among the the experimental group (PRF), the positive control group (bone meal group) and the negative control group (blank group).1-3 days after teeth extraction, the mucosa of the positive control groups and negative control groups appeared slight red,while the experimental group 's was normal. The surface of extraction wounds were completely covered by epithelial tissue after five days .
The relative length of the remaining alveolar ridge observed by X-ray resulted as follows: the experimental group (PRF)> the positive control group (bone meal group) >the negative control group (control group), the difference was statistically significant ( P <0.05) after 1 week, 2 weeks and 4 weeks. After 8 weeks, 12 weeks, the experimental group (PRF) and the positive control group (bone meal group) are greater than the negative control group (control group)(P<0.05) .But there was no significant difference between the experi- mental group (PRF) and the positive control group (bone meal group)( P >0.05).
The quantity and quality of new bone in the sockets of the experimental group (PRF ) were better than the positive control group (bone meal) and negative control group in the histological observation of 1 week, 2 weeks and 4 weeks . There was a large number of new bone along the nest wall which grew to the center of the sockets in the experimental group in 1 week and the experi- mental group had more lacunae and active osteoblasts than the positive control and the negative control group.There was still new bone formed in the positive control group (bone meal group) after 8 and 12 weeks,however,of which the experimental group (PRF group) and positive control group (bone meal group) had been completed remodelled, there was no significant difference on the extent of calcification in the bone lamellar structure between the two groups.
Conclusion
In the experimental group,we compared the restoration of the remaining alveolar bone by implanting the PRF ,hydroxyapatite in the sockets and the blank control after teeth extraction.All in all,the conclusions were as follows:
1、PRF and hydroxyapatite bioceramics (bone meal) can all preserve the height of the alveolar ridge after teeth extraction while PRF is more evident at an early stage.
2、PRF can restore the alveolar ridge as a independent transplant material.
34、PRF can remedy the inflammatory symptoms which can reduce post-traumatic inflammation and accelerate the healing process of wound.
引文
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[7] Man D,PloskerH,Winland-Brown JE.The use of autologous platelet-rich plasma(platelet gel)and autologous platelet poor plasma(fibrin glue)in cosm- etic surgery[J].Plast Reconstr Surg,2001,107(1):229-237.
[8] Choukroun J, Adda F, Schoeffler C, et al. Une opportunite′en paro- implantologie: le PRF. Implantodontie 2000;42:55-62.French
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[15] Cohen S.The stimulation of epidermal proliferation by a Specific protein (EGF) [J].Dev Bid,1965,12:394-407.
[16] Schmidt MB,Chen EH,Lynch SE.A review of the effects of insulin-like growth factor and platelet derived growth factor on in vivo cartilage healingand repair[J].Osteoarthritis&Cartilage,2006,14(5):403-412.
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[18]周延民,李琦,储顺礼等.即刻种植间隙充填富血小板纤维蛋白修复骨缺损2例[J].中国实用口腔科杂志,2009, 2(8)504-505.
[19]李琦.Choukroun富血小板纤维蛋白修复兔颅骨骨缺损的实验研究[D].吉林大学,2009.
[20]吴哲.载辛伐他汀聚合物对剩余牙槽嵴骨修复影响的实验研究[D].吉林大学,2007.
[21] Carnin AN,Simons A,Klein M,et al.The use of a Partiuclate micor Poorus, calcified copolymer as a ridge maintenance device in dog[J]. VetDent. 1995, 12 (2): 53-8.
[22] Murata M,Huang BZ,Shibata T,et al.Bone augmentation by recombinant human BMP-2 and collagen on adult rat parietal bone[J].Int J Oral Maxi- llofac Surg,1999,28:232-237.
[23] Kamakura S,Sasano Y,Nakamura M,et al.Initiation of alveolar ridge augmentation in the rat mandible by subperiosteal implantation of octacalc- ium phosphate [J]. Archs Oral Biol,1996.41(11):1029-1038.
[24]吴哲,孙宏晨,车彦海等.大鼠下颌切牙拔出后剩余牙槽嵴吸收模型的建立[J].实用口腔医学杂志,2007,23(1)36-39.
[25] Choukroun J,Diss A,Simonpieri A,et al.Platelet-rich fibrin (PRF):a second- generation platelet concentrate.Part IV: clinical effects on tissue healing [J].Oral Surgery Oral Medicine Oral Pathology Oral Radiology &End- odontics,2006,101(3):e56-e60.
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[2] Van Sickels JE,Montgomeyr MT.Review of sugrical ridge augmentation porcedures for the atrophied mandible[J]. J Porsthet Dent, 1984: 51(1):5-10.
[3]付小兵,王德文.创伤修复基础[M],第1版.北京:人民军医出版社,1997:129-204.
[4] Assoian RK,Grotendorst GR,Miller DM, et al.Cellular transformation on by coordinated action of three peptide growth factors from human platelets [J].Nature,1984,309(5971):804-806.
[5] Marx RE,Carlson ER,Eichstacdt RM, et al.Platelet-rich Plasma; Growth factor enhancement for bone grafts[J].Oral Surgery Oral Medicine Oral Pathology Oral Radiology &Endodontics,1998,85(6):638-656.
[6] Wrotniak M,Bielecki T,Gazdzik TS.Current opinion about using the platelet- rich gel in orthopaedics and trauma surgery[J].Ortopedia Traumatologia Rehabilitacja,2007,9(3):227-238.
[7] Man D,PloskerH,Winland-Brown JE.The use of autologous platelet-rich plasma(platelet gel)and autologous platelet poor plasma(fibrin glue)in cosm- etic surgery[J].Plast Reconstr Surg,2001,107(1):229-237.
[8] Choukroun J, Adda F, Schoeffler C, et al. Une opportunite′en paro- implantologie: le PRF. Implantodontie 2000;42:55-62.French
[9] Nowzari,Matian,Slots.Periodontalpathogens on Polytetrafluoroethylene me- mbranes for guided tissuee generation inhibithealing[J].Journal of ClinicalPeriodontology,1995,22:469-474.
[10] 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 Surgery Oral Medicine Oral Pathology Oral Radiology &Endodontics,2006,101:e51-e55.
[11] Badiavas EV, Abedi M, Butmarc J, et al. Participation of bone marrow derived cells in cutaneous wound healing[J]. Cell Physiol 2003,196: 245-50.
[12] Choukroun J,Diss A,Simonpieri A,et al.Platelet-rich fibrin (PRF):a second- generation platelet concentrate.Part IV: clinical effects on tissue healing [J].Oral Surgery Oral Medicine Oral Pathology Oral Radiology & Endodontics. 2006;101(3):e56-60.
[13] Sporn MB,Roberts AB,Wakefield LM, et al.Transforming growth factor- beta:biological function and chemical [J].Science, 1986, 233 (4763): 532- 534.
[14] Lucarelli E,Beccheroni A,Donati D.Platelet-derived growth factors enhance proliferation of human stromal stem cells[J]. Biomaterials, 2003, 24(11): 3095-3100.
[15] Cohen S.The stimulation of epidermal proliferation by a Specific protein (EGF) [J].Dev Bid,1965,12:394-407.
[16] Schmidt MB,Chen EH,Lynch SE.A review of the effects of insulin-like growth factor and platelet derived growth factor on in vivo cartilage healingand repair[J].Osteoarthritis&Cartilage,2006,14(5):403-412.
[17] Choukroun J,Diss A,Simonpieri A, et al.Platelet-rich fibrin(PRF):A second- generation platelet concentrate.PartV:Histologic evaluations of Choukroun’s PRF effects on bone allograft maturation in sinus lift[J].Oral Surgery Oral Medicine Oral Pathology Oral Radiology &Endodontics, 2006,101: 299- 303.
[18]周延民,李琦,储顺礼等.即刻种植间隙充填富血小板纤维蛋白修复骨缺损2例[J].中国实用口腔科杂志,2009, 2(8)504-505.
[19]李琦.Choukroun富血小板纤维蛋白修复兔颅骨骨缺损的实验研究[D].吉林大学,2009.
[20]吴哲.载辛伐他汀聚合物对剩余牙槽嵴骨修复影响的实验研究[D].吉林大学,2007.
[21] Carnin AN,Simons A,Klein M,et al.The use of a Partiuclate micor Poorus, calcified copolymer as a ridge maintenance device in dog[J]. VetDent. 1995, 12 (2): 53-8.
[22] Murata M,Huang BZ,Shibata T,et al.Bone augmentation by recombinant human BMP-2 and collagen on adult rat parietal bone[J].Int J Oral Maxi- llofac Surg,1999,28:232-237.
[23] Kamakura S,Sasano Y,Nakamura M,et al.Initiation of alveolar ridge augmentation in the rat mandible by subperiosteal implantation of octacalc- ium phosphate [J]. Archs Oral Biol,1996.41(11):1029-1038.
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