纤维蛋白胶复合物促进犬骨折愈合的临床学观察

为了探索纤维蛋白胶(FG)联合人骨形态发生蛋白(rhBMP-2)、碱性成纤维细胞生长因子(bFGF)和妥布霉素的复合物对犬骨折愈合的影响,建立了犬胫骨骨折模型,将FG复合物应用于实验组和空白对照组的胫骨骨折处,在术后2、4、8、12、16周,测定骨折处周围软组织充血水肿消除时间、利用X射线影像学评价骨折愈合程度、测定骨折愈合部位的骨密度(BMD)及骨密度比率、骨痂体积,测定胫骨生物力学强度并进行统计和分析。实验组和对照组比较:术部红、肿、热消失时间显著缩短,术后各时间点的X射线影像学评分更高,术后各时间点骨痂更大,术后16周术部骨骼能承受的最大载荷和最大位移显著增大。结果表明FG联合rhBMP-2、bFGF与妥布霉素应用于骨折部位,可促进骨折愈合过程中软组织的生成,缩短骨折愈合的时间,提高骨生物力学强度。     

纤维蛋白粘合剂在犬小肠端端吻合术中对吻合口Hyp、VEGF与bFGF含量的影响

为探讨纤维蛋白粘合剂(FG)在犬小肠端端吻合术中对吻合口处羟脯氨酸(Hyp)、血管内皮生长因子(VEGF)与碱性成纤维细胞生长因子(bFGF)含量的影响,将30只试验犬随机分为试验组和对照组两组,试验组采用简易缝合配合纤维蛋白粘合剂,对照组采用常规手术肠线缝合;分别于术后第3、5、7、14和28天每组随机选取3只测定吻合口处Hyp、VEGF与bFGF的含量。术后试验组Hyp含量在第5天显著高于对照组(P0.05),bFGF的含量在第3天、第5天、第7天显著高于对照组(P0.05),VEGF含量在第3天和第5天高于对照组(P0.01),在第14天和第28天显著低于对照组(P0.01)。结果表明,应用简易缝合配合纤维蛋白粘合剂吻合肠管,可促进吻合口处小肠组织内羟脯氨酸、血管内皮因子和碱性成纤维细胞生长因子的分泌,从而加速吻合口的愈合。     

碱性成纤维细胞生长因子对犬穿透性角膜移植愈合效果的组织学观察

为探索碱性成纤维细胞生长因子(bFGF)对犬穿透性角膜移植(PKP)术后角膜愈合效果,选取24只健康犬接受角膜移植,随机分为4组:手术对照组(A)、地塞米松组(B)、地塞米松+环孢菌素A组(C)、地塞米松+环孢菌素A+bFGF组(D),所有犬术后均接受妥布霉素点眼,在术前(0 d)和术后14 d、35 d制作角膜组织切片,观察角膜结构变化。结果显示:A组犬角膜基质层纤维排列混乱,上皮恢复速度较慢,35 d时存在上皮与基质层分离现象;B、C组角膜基质层纤维排列不规则,上皮细胞完整覆盖创面;D组角膜基质层纤维排列整齐,上皮层恢复速度最快,角膜表面平滑。结果表明:bFGF与妥布霉素、地塞米松和环孢菌素A联用可促进PKP术后角膜上皮层和基质层的愈合,为临床上犬角膜移植的术后治疗提供参考。     

移植脐带间充质干细胞对荷斯坦公犊日增重及血清生长因子水平的影响

本试验旨在研究移植脐带间充质干细胞对荷斯坦公犊日增重及血清生长因子水平的影响.选取1月龄体重相近的荷斯坦公犊28头,随机分为对照组和试验组,采集健康的荷斯坦新生胎牛的脐带组织,通过胰酶消化筛选法体外分离、纯化及培养牛脐带间充质干细胞(UC-MSCs),试验组按公犊体重(3×10⁵个/kg干细胞)稀释至8 mL颈静脉注射至体内,对照组注射等量生理盐水.试验期154 d.结果表明,注射UC-MSCs后,试验组公犊平均体重在4、5、6月龄时显著高于对照组(P< 0.05);试验组公犊平均日增重在3、4月龄时显著高于对照组(P< 0.05),而在6月龄时极显著高于对照组(P< 0.01).试验组血清中类胰岛素样生长因子(IGF-1)水平在4月龄时显著高于对照组(P< 0.05);试验组血清中血管内皮生长因子(VEGF)、转化生长因子(TGF-α)水平在3月龄时显著高于对照组(P< 0.05);试验组血清中VEGF、成纤维细胞生长因子(bFGF)、TGF-α水平在6月龄时显著或极显著高于对照组(P< 0.05;P< 0.01);2、3、4、6月龄时,试验组血清中骨形态发生蛋白(BMP-7)水平显著低于对照组(P< 0.05).综上所述,移植UC-MSCs能显著提高公犊的平均日增重及血清中IGF-1、VEGF、TGF-α、bFGF水平,血清中这些生长因子与公犊的生长发育密切相关,进而影响其生长性能.     

纤维蛋白胶的止血效果观察

在6只犬的肝和脾分别作长1．5和0.5cm深的切口，在不阻断肝，脾血液供应等措施下，分别施用和不施用纤维蛋白胶作为试验组和对照组处理创口，对此其出血量及止血时间，来观察纤维蛋白胶对犬肝，脾实验性损伤的止血效果。结果表明，试验组肝切口的出血量（0.038mL）和止血时间（11s）较对照组（0.230mL,102s）明显减少，试验组脾切口的出血量（0.050mL)和止血时间（15.7s)较对照组（0.330mL,199s)显著减少，术后7d和14d再次开腹检查，发现试验组的肝，脾伤口愈合良好，未见组织粘连，腹腔感染等现象。     

N-BCA医用胶在犬眼睑手术的应用研究

为了评估氰基丙烯酸正丁酯(N-BCA)医用胶在犬眼睑手术的安全性和临床效果,对12只健康犬的24只眼进行试验。将双眼分为2组:左眼使用N-BCA医用胶闭合眼睑创口(试验组),右眼使用传统缝合材料闭合眼睑创口(对照组)。术后进行常规眼科检查,观察并记录眼睑创口闭合时间,犬术后整体状态,以及创口愈合时间。术后7 d、14 d和28 d,取眼睑皮肤进行显微镜下组织病理学观察。结果显示,术后试验组和对照组眼睑皮肤创口愈合良好。创口闭合时间和温哥华瘢痕量表评分在试验组和对照组之间有显著性差异(P0.05)。在眼睑皮肤样本组织病理学观察中,对照组瘢痕更明显,有成纤维细胞增生。试验表明,氰基丙烯酸正丁酯医用胶的使用是一种有效的闭合犬眼睑皮肤创口的方法,对眼睑皮肤组织和眼表无明显影响,可用于犬眼睑手术。     

一例犬右侧前后肢同时骨折内固定手术治疗

本文结合1例犬肱骨远端骨折、胫骨中端骨折内固定手术的过程,介绍了这个病例从术前、术中到术后的具体情况。结果表明,该犬采用塑形钢板加柯氏针对肱骨远端骨折进行固定和采用DCP接骨板对胫骨中端骨折内固定整复后效果良好,术后第4d该犬就能正常下地行走,2周后基本恢复正常。通过这个病例的研究,证明了采用内固定的手术疗法可使骨折很快愈合,治疗效果也能达到预期目的。     

一剂复方中药对家兔骨骼愈合的影响

复方中药对骨折愈合的作用机制目前尚不明确,本试验通过观察复方中药对家兔桡骨骨折后血清钙、磷、碱性磷酸酶和X线影像学等指标的影响,分析该中药促骨愈合的效能及其可能作用机理。试验选用48只成年家兔,制成骨缺损标准模型,随机分为试验组和空白对照组,试验组每日在饲料中添加复方中药2.5 g/kg饲喂,对照组给予普通饲料。分别于术后2、4、6、8周进行生化指标和X线影像学检测。试验结果表明,试验组血清钙、血清磷、血清碱性磷酸酶及X线片评分骨折后明显高于对照组。该复方中药能缩短骨折愈合过程所需时间,提高骨折愈合质量,有利于骨折愈合。     

犬硬膜外阻滞对腹部手术应激的初步研究

8例犬间隔1个月分别施行2次腹部手术,第1次采用硬膜外阻滞(试验组,n=8),第2次采用硫喷妥钠静脉麻醉(对照组,n=8)。分别于麻醉前(T0)、麻醉后(T1)、切开腹壁时(T2)、手术30min(T3)、手术完毕时(T4)、手术后24h(T5)、手术后48h(T6)7个时间静脉采血,采用放射免疫法测定血浆皮质醇、白细胞介素-2浓度,采用比色法测定血糖浓度。结果:(1)对照组血浆皮质醇浓度平均值显著高于试验组(P<0·05),尤其在T3、T4时显著高于试验组(P<0·05)。对照组在T3、T4时血浆皮质醇浓度显著高于T0(P<0·05),而试验组各时间与T0比较均无显著差异(P>0·05);(2)对照组血浆白细胞介素-2浓度平均值极显著低于试验组(P<0·01),尤其在T1、T2、T5和T6时显著低于试验组。对照组在T5时、试验组在T3、T4时其血浆白细胞介素-2浓度均显著低于T0;(3)对照组血糖浓度平均值显著高于试验组,尤其在T4时显著高于试验组。对照组在T3、T4、T5和T6时、试验组则在T4时其血糖浓度均显著高于T0。结论:硬膜外阻滞组与硫喷妥钠静脉麻醉比较,硬膜外阻滞能够更有效缓解犬腹部手术的应激反应。     

乳香提取物对大鼠皮肤周围神经损伤修复影响

通过乳香提取物(frankincense extract)在大鼠皮肤缺损模型中对周围神经损伤修复的影响,探讨乳香提取物对周围神经损伤的修复作用。将25只SD大鼠造皮肤缺损模型,采用自身对照。一侧为对照组,每日常规消毒;另一侧为试验组,每日涂抹乳香提取物。分别于1、4、7、10、14d5个时间点对创面进行拍照,测量创口面积,并取5只大鼠的创面皮缘备用。用免疫组织化学法检测S-100蛋白的表达及RT-PCR法检测碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)、神经生长因子(nerve growth factor,NGF)基因表达量的变化。试验结果表明,创伤愈合率显示,试验组较对照组伤口能更快愈合,且试验组第14天伤口基本愈合。免疫组织化学显示,S-100蛋白试验组较对照组有更多的阳性表达,且在第7天阳性结果更明显。RT-PCR显示,试验组bFGFmRNA表达较对照组有增多趋势,第4和7天与对照组相比差异显著(P<0.01)。试验组NGFmRNA表达较对照组相比,差异均显著(P<0.01)。第7和10天NGFmRNA表达多于对照组。     

Effect of Gelatin Hydrogel Sheet Containing Basic Fibroblast Growth Factor on Proximal Sesamoid Bone Transverse Fracture Healing in the Horse

Our objective in this study was to examine the effect of gelatin hydrogel (GH) sheets containing basic fibroblast growth factor (bFGF) on healing of proximal sesamoid bone transverse fractures in the horse. Ten healthy adult Thoroughbreds were used. The lateral proximal sesamoid bone of the left forelimb and the medial proximal sesamoid bone of the right forelimb were osteotomized, while the horses were under general anesthesia, and subsequently repaired by lag screw fixation using a single 4.5-mm cortical screw. A GH sheet containing 100 μg of bFGF was then sutured to the synovial membrane adjacent to the osteotomized proximal sesamoid bone. In the control group, the fracture was fixed with a lag screw, and the articular capsule was sutured. Fracture healing was assessed by radiographic examination once a week for 16 weeks after the operation. Radiographic examination of bone healing revealed significantly lower demineralization of the fracture line in the GH sheet-treated group than in the control group. The rate of demineralization of the fracture line in the GH sheet-treated group was significantly lower than that in the control group at 2, 4, and 8 weeks after the operation. In this study, we demonstrated that the use of a GH sheet containing bFGF promotes healing of proximal sesamoid bone fracture in the horse. Therefore, it is believed that this treatment strategy would be useful for quick recovery from bone fracture in the horse.     

Recombinant human bone morphogenetic protein-2 in absorbable collagen sponge enhances bone healing of tibial osteotomies in dogs

OBJECTIVE: To compare the efficacy of 2 doses of recombinant human bone morphogenetic protein-2 (rhBMP-2) on tibial osteotomy healing in dogs. STUDY DESIGN: Experimental, randomized complete block (n=7). ANIMALS: Adult female dogs (n=21). METHODS: Right midshaft tibial osteotomies were created and stabilized with a 1-mm gap using type I external fixators. Seven dogs were untreated controls and 14 with osteotomies were treated with either 0.05 or 0.2 mg/mL rhBMP-2 delivered in an absorbable collagen sponge (ACS). At 8 weeks, dogs were euthanatized and bones were mechanically tested and examined by microscopy. RESULTS: Bone healing based on radiographic scoring, was significantly improved in dogs treated with 0.2 mg/mL of rhBMP-2 compared with the other groups; these tibiae were also significantly stronger and stiffer than 0.05 mg/mL rhBMP-2 and control osteotomized tibiae. Histologic scores were significantly better for 0.2 mg/mL rhBMP-2 group than 0.05 mg/mL rhBMP-2 group, but neither was significantly different from control. CONCLUSIONS: rhBMP-2 in ACS at a concentration of 0.2 mg/mL improves healing of tibial osteotomies in dogs compared with untreated controls and 0.05 mg/mL rhBMP-2 based on force plate analysis and radiographic evaluation. This was not confirmed histologically but treated bones had improved mechanical properties at 8 weeks. CLINICAL RELEVANCE: After a long bone fracture, dogs may face a long recovery period before full return of limb function. rhBMP-2, in association with good fracture fixation principles, may enhance bone healing in dogs with diaphyseal fractures.     

Comparison of two doses of recombinant human bone morphogenetic protein in absorbable collagen sponges for bone healing in dogs

OBJECTIVE: To determine the effects of 2 doses of recombinant human bone morphogenetic protein-2 in an absorbable collagen sponge (rhBMP-2/ACS) on bone healing in dogs. ANIMALS: 27 adult dogs. PROCEDURES: Dogs underwent a mid-diaphyseal (1-mm) tibial osteotomy (stabilized with external skeletal fixation) and received an ACS containing 0.28 mg (0.2 mg/mL) or 0.56 mg (0.4 mg/mL) of rhBMP-2 or no treatment (control dogs). All dogs were examined daily; bone healing was assessed via radiography and subjective lameness evaluation every 2 weeks. After euthanasia at 8 weeks, tibiae were evaluated biomechanically and histologically. RESULTS: Control dogs required antimicrobial treatment for pin-site-related complications more frequently than did rhBMP-2/ACS-treated dogs. At 4 and 6 weeks, weight bearing was greater in dogs treated with rhBMP-2/ACS (0.2 mg/mL) than in control dogs, albeit not significantly. Compared with control treatment, both doses of rhBMP-2/ACS accelerated osteotomy healing at 4, 6, and 8 weeks, and the 0.2 mg/mL dose enhanced healing at 2 weeks; healing at 6 weeks was greater for the lower-dose treatment than for the higher-dose treatment. Histologically, healing at 8 weeks was significantly improved for both rhBMP-2/ACS treatments, compared with control treatment. Among groups, biomechanical variables did not differ, although less osteotomy-site failures occurred in rhBMP-2/ACS-treated groups. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs that underwent tibial osteotomy, rhBMP-2/ACS (0.2 mg/mL) appeared to accelerate bone healing and reduce lameness (compared with control treatment) and apparently augmented bone healing more than rhBMP-2/ACS (0.4 mg/mL). Compared with control dogs, rhBMP-2/ACS-treated dogs required antimicrobial treatments less frequently.     

Clinical Use of Recombinant Human Bone Morphogenic Protein-2 in a Whooping Crane (Grus americana)

Objective— To report use of recombinant human bone morphogenetic protein-2 (rhBMP-2) as adjunctive therapy for treatment of a comminuted, open, proximal humeral fracture in an avian species.
Study Design— Clinical report.
Animals— A 3.5-month-old male whooping crane ( Grus americana ).
Methods— An open, severely comminuted humeral facture was stabilized with an intramuscular (IM) pin/type IA external skeletal fixator with tie-in configuration. rhBMP-2 was applied in a calcium phosphate matrix (CPM) paste directly to the fracture site as a bone graft substitute. Radiographic evidence of bone healing was monitored for 14 weeks.
Results— Substantial bony callus was evident at 4 weeks and at 8 weeks there was bridging callus with obvious bony remodeling. The fixation was destabilized at 9 weeks by IM pin removal, bone healing progressed and the fixator was removed at 11 weeks. By 14 weeks both cortices had been re-established with continued callus remodeling evident.
Conclusion— rhBMP-2, applied in a CPM paste, was used as a bone graft substitute in the treatment of a comminuted, open humeral fracture in a whooping crane.
Clinical Relevance— Use of rhBMP-2/CPM should be considered in treatment of avian fractures.     

Comparison of fibrin glue and suture in the healing of teat incisions in lactating goats

The aims of this study were to investigate whether fibrin glue can be used to close experimentally induced incisions of the teat (mammary papillae) in lactating goats and to compare the healing of the glued with the sutured incisions. Four clinically healthy lactating dairy goats, namely 8 mammary papillae were used. After surgical preparation of the papillae, a 3.5 cm long incision of each papilla was made through skin, muscular layer and mucosa into the papillary sinus. The wounds in the right papillae in all goats were closed with U-shaped uninterrupted 00 chromic catgut sutures. The wounds in the left papillae in all goats were closed, using fibrin glue. One incision was seen to be dehisced and fistulous one day after in fibrin glued teats. The animals were slaughtered 8 days after surgical manipulation. The mammary papillae were removed and examined in the viewpoint on gross and microscopic findings. The healing of wounds was slower and feeble in glued mammary papillary incisions, however faster and stronger in sutured incisions on day 8 after operations. But, available outcomes like less tissue thickness and positive cosmetic results could be obtained byfibrin glue used on mammary papillary incisions, which are very important for teats to be milked by hand and milking machine. Results suggest that it is advisable to use only one or two simple interrupted sutures in teat incisions glued with fibrin to prevent the dehiscence but with a more reliable healing than the sutured incisions.     

Histology and immunohistochemistry study of ovine tendon grafted with cBMSCs and BMMNCs after collagenase-induced tendinitis

OBJECTIVES: The aim of this study was to compare the regeneration abilities of cultured bone marrow mesenchymal cells (cBMSC) and bone marrow mononuclear cells (BMMNC) with fibrin glue, saline solution and sham control in collagenase-induced tendinitis of the Achilles tendon in sheep. METHODS: Six sheep were recruited randomly to each group: cBMSC, BMMNC, fibrin, saline and sham control. Each group received the relative treatment two weeks after inducing lesions (T(0)). After eight weeks (T(8)) of treatment, the tendons were harvested and evaluated for histomorphology, Collagen type I, III, Cartilage Oligomeric Matrix Protein (COMP) and CD34 positive cells expression. RESULTS: Histology and immunohistochemistry showed similar capabilities of cBMSC and BMMNC to restore the architecture of fibres and Extra Cellular Matrix (ECM), with a high expression of collagen type I and COMP and a very low expression of collagen type III in treated tendons. The complete architectural disruption of fibres, dramatic reduction of collagen Type I and COMP expression and increase collagen type III expression were commonly observed in tendons treated with fibrin or saline only. The presence of CD34 positive cells was appreciable in the BMMNC group while few cBMSC showed this cluster of differentiation, not expressed in tendons treated with fibrin or saline. CLINICAL SIGNIFICANCE: The data in this study show the efficacy of cBMSC and BMMNC in regenerating tendon tissue after collagenase-induced tendinitis.     

Effects of isobutyl-2-cyanoacrylate on bone healing

An in vivo evaluation of isobutyl-2-cyanoacrylate as an osseous adhesive was performed to determine its effect on bone healing. Oblique cortical fractures were induced at the tibial-fibular junctions of adult rabbits. Forty-eight fractures were repaired by the use of isobutyl-2-cyanoacrylate and a full cerclage wire. The opposite control fracture was repaired with a full cerclage wire only. One hundred percent of the glued fractures were not united through the glue interface, although normal periosteal new bone formation (secondary bone healing) took place around the glue. One hundred percent of the nonglued fractures healed by secondary bone union. Reactions were not seen in the bone or soft tissue adjacent to the adhesive material.     

Basic fibroblast growth factor stimulates epithelial cell growth and epithelial wound healing in canine corneas

Objective  To evaluate the effect of basic fibroblast growth factor (bFGF) on the proliferation of canine corneal epithelial cells and epithelial wound healing.
Animal studied  Canine corneal epithelial cells from the corneas of euthanized dogs and corneal epithelial wounds on one eye from each of 24 dogs.
Procedures  The proliferation of corneal epithelial cells in vitro was measured using the methylthiazolyl-tetrazolium (MTT) assay. A corneal wound on one eye of each dog was made with a corneal trephine (6 mm diameter). Four concentrations of bFGF, 0, 100, 500, and 1000 ng/mL, were applied to the affected eyes of dogs, t.i.d. Fluorescein staining was used to assess closure of the corneal epithelial wound.
Results  The addition of bFGF resulted in a significant increase in epithelial proliferation at 24 h after culture, except 1 ng/mL bFGF. Cells with all bFGF treatments proliferated significantly at 48 and 96 h compared to those in the non-bFGF group. bFGF at a concentration of 10 ng/mL promoted cell proliferation maximally. The wound healing rate in the bFGF-treated groups was greater than that in the control. All corneal wounds in bFGF-treated corneas closed by day 7, whereas two of six corneal wounds in the control showed poor healing. None of the eyes developed corneal clouding or neovascularization during the experiment.
Conclusions  Basic fibroblast growth factor accelerated the proliferation of canine epithelial cells and effectively promoted corneal epithelial wound healing.     

Treatment of nonunions with nonglycosylated recombinant human bone morphogenetic protein-2 delivered from a fibrin matrix

OBJECTIVE: To report the results of the treatment of nonunions with nonglycosylated recombinant human bone morphogenetic protein-2 (nglBMP-2) delivered from a designed fibrin matrix. STUDY DESIGN: Experimental trial in rodents and prospective clinical study in dogs and cats with nonunion fractures. ANIMALS: Twenty adult female, albino, Sprague-Dawley rats; 8 client-owned cats and dogs. METHODS: After development of a fibrin matrix and evaluation of nglBMP-2 in a rodent femoral defect model, 8 consecutive long bone nonunion fractures (no progression in healing in > or = 3 months), were treated using 300 microg nglBMP-2 in a liquid fibrin precursor, injected into the defect gap after fracture revision and stabilization, or through a stab incision into the fracture site. The fibrin matrix was designed to clot in the wound after 60 seconds and to release the nglBMP-2 continuously over several days. RESULTS: Using only fibrin gel, 7% of the rat femoral defect was filled with new formed bone compared with 79% defect filling using 2 microg nglBMP-2 (P=.006). Five and 10 microg nglBMP in fibrin resulted in union of all femoral defects with complete filling of the gap with new bone. Bony bridging and clinical healing was achieved in 7 patients within 24 weeks of administration of nglBMP-2. CONCLUSIONS: Application of nglBMP-2 in a functional matrix can induce bone healing. Controlled release of nglBMP-2 from a fibrin matrix mimics the natural fracture hematoma. CLINICAL RELEVANCE: nglBMP-2/fibrin can successfully replace a cancellous bone autograft in fracture treatment with an associated reduction in graft donor site morbidity and surgical time.