Tissue Regeneration in Infected Wounds of Albino Rats Using Ciprofloxacin-Loaded Gelatin Microspheres Incorporated Collagen Scaffold: A Histological Approach with H&E Staining

A wound care system consisting of ciprofloxacin-loaded gelatin microspheres impregnated in a macroporous collagen scaffold was created to effectively control wound infection and regenerate soft tissue at the wound site.Histological and biochemical alterations were observed in infected wounds treated with these scaffolds in Albino Wistar rats.Furthermore,the study examined the immediate and prolonged release of ciprofloxacin from the scaffolds,as well as their function in eliminating bacterial infections and expediting the process of skin healing and regeneration.The developed technique was followed in the streamlined process of creating these collagen scaffolds.Compared to untreated wounds,the group receiving scaffold treatment experienced a faster rate of wound closure.It was noted that the rate of infections was considerably reduced and that full soft tissue regeneration occurred within 12 days.The development of well-deposited collagen bundles in the treated groups was demonstrated by H&E staining,which verified the flawless regeneration of the dermis and epidermis.The antimicrobial agent-loaded gelatin microspheres impregnated into the porous collagen scaffold demonstrated remarkable soft tissue regeneration and efficient infection control at the wound site.

Studies on Adriamycin Magnetic Gelatin Microspheres

The preparation and properties of adriamycin magnetic gelatin microspheres(Adr- MG-ms)were reported.The synthesis of magnetic iron oxide ultrafine particle and embolization effects of magnetic gelatin microspheres(MG-ms)in dog were studied.Adr- MG-ms consist of 2%(w/w)of adriamycin(Adr)as the core,and 68% of gelatin and 30% of magnetite as the shell with a mean particle size of 22 μm. In vitro experiment,the release rate of drug demonstrated that the microspheres have sustained-release properties.The average diameter of magnetic iron oxide was approximately l0 nm. Transcatheter embolization with MG-ms and ￣(99m)Tc-labelled MG-ms was performed under external magenet control in dog liver,respectively.Gamma photography and angiogram revealed that MG-ms level was almost equal both in left and right hepatic arteries without magnet,while with magnet(1200 Gs),MG-ms level in left hepatic artery(target site)was about 2.25 fold higher than in right hepatic artery,and few MG-ms in thyroid gland,brain and heart was observed.Results showed that the MG-ms is a promising embolic agent for treatment of hepatic cancer under external magnet control.

Preparation and Properties of Gelatin-Chitosan/Montmorillonite Drug-loaded Microspheres

A kind of slow release drug-loaded microspheres were prepared with gelatin, chitosan and montmorillonite（MMT） by an emulsification/chemical cross-linking method using glutaraldehyde as cross-linking agent and acyclovir as model drug. The microspheres were characterized by X-ray diffraction （XRD）, Fourier transform infrared （FT-IR） and scanning electron microscopy （SEM）, respectively. The morphology, drug content, encapsulation efficiency and drug-release behavior were investigated with different MMT contents. The experimental results indicated that intercalated microspheres could be prepared, the morphology of microspheres was markedly affected by MMT. The glomeration performance of uncross-linked microspheres was improved because of the physical cross-linking of MMT. Drug content and encapsulation efficiency were decreased when increased the content of MMT, but burst release and the drug release were significantly decreased with the addition of MMT. Effective physical cross-linking could be formed when added MMT, and MMT could reduce the content of toxic chemical cross-linking agents.

Preparation and in vitro Release Performance of Sustained-release Captopril/Chitosan-gelatin Net-polymer Microspheres

The captopril/Chitosan-gelatin net-polymer microspheres(CTP/CGNPMs) were prepared using Chitosan(CTS) and gelatin(GT) by the methods of emulsification,cross-linked reagent alone or in combination and microcrystalline cellulose(MCC) added in the process of preparation of microspheres,which aimed to eliminate dose dumping and burst phenomenon of microspheres for the improvement of the therapeutic efficiency and the decrease of the side effects of captopril(CTP). The results indicated that CTP/CGNPMs had a spherical shape,smooth surface and integral structure inside but no adhesive phenomena in the preparation. The size distribution ranged from 220 μm to 280 μm. The CTP release test in vitro demonstrated that CTP/CGNPMs played the role of retarding the release of CTP compared with ordinary CTP tablets. The release behaviors of CGNPMS were influenced by preparation conditions such as experimental material ratio(EMR) and composition of cross linking reagents. Among these factors,the EMR(1/4),CLR(FA+SPP) and 0.75% microcrystalline cellulose(MCC) added to the microspheres constituted the optimal scheme for the preparation of CTP/CGNPMs. The ER,DL and SR of CTP/CGNPMs prepared according to the optimal scheme were 46.23±4.51%,9.95±0.77% and 261±42%,respectively. The CTP/CGNPMs had the good characteristics of sustained release of drug and the process of emulsification and cross-linking were simple and stable. The CGNPMs are likely to be an ideal sustained release formulation for water-soluble drugs.

Preparation and Characterization in vitro of Sustained-release Captopril/ Chitosan-gelatin Net-polymer Microspheres(Cap/CGNPMs)

The captopril/ Chitosan-gelatin net-polymer microspheres （ Gap/ CGNPMs ） were prepared using Chitosan （ CS ） and gelatin （ Gel ） by the methods of emulsification. A cross linked reagent alone or in combination with microcrystalline cellulose （ MCC ） was added in the process of preparation of microspheres to eliminate dose dumping and burst phenomenon of microspheres for the improvemeat of the therapeutic efficiency and the decrease of the side effects of captopril （ Cap ）. The results indicate that Cap/ CGNPMs have a spherical shape , smooth surface roorphology and integral inside structure and no adhesive phenomena and good roobility , and the size distribution is mairdy from 220 to 280 μm. Researches on the Cap release test in vitro demonstrate that Cap/ CGNPMs are of the role of retarding release of Cap compared with Cap ordinary tablets （COT）, embedding ratio （ER） , drug loading （ DL ）, and swelling ratio （ SR ）, and release behaviors of CGNPMS are influenced by process conditions of preparation such as experimental material ratio （EMR） , composition of cross linking reagents. Among these factors , the EMR（1/4）, CLR （ FOR ＋ TPP） and 0.75% microcrystulline cellulose （MCC） added to the microspheres are the optimal scheme to the preparation of Cap/CGNPMs. The Cap/CGNPMs have a good characteristic of sustained release of drug, and the process of emulsifieation and crossinking process is simple and stable. The CGNPMs is probable to be one of an ideal sustained release system for water-soluble drugs.

Preparation, Characterization, and in vitro Release of Biodegradable Erythromycin-gelatin Microspheres

Blank and erythromycin-loaded gelatin microspheres were successfully fabricated via emulsion chemical- crosslinking technique. The surface morphology of the microspheres was characterized by scanning electron microscope（SEM） and optical microscope. The results show that the microspheres were spherical and smooth. The particle average size of erythromycin-loaded microspheres was found to be 20.6 μm, with a high purity of more than 90% and with a good dispersibility. The microspheres could be obtained in a high yield. Erythromycin released from the microspheres was monitored in buffer and artificial body fluid at 37 ℃. Average drug content was 27.2%, and erythromycin-loaded gelatin microspheres showed good release profiles with a nearly constant release during 4-8 h in artificial body fluid in vitro degradation studies. These gelatin microspheres are useful for studying and developing various drug-delivery systems.

PREPARATION AND DRUG RELEASE CHARACTERISTICS OF PINGYANGMYCIN GELATIN MICROSPHERES FOR EMBOLIZATION THERAPY

Objective: To prepare Pingyangmycin gelatin microspheres (PYM-GMS) for carotid artery embolization therapy and to study the release characteristics in vivo and in vitro. Methods: PYM-GMS was prepared by optical double-phase emulsified condensation polymerization. Through UV-spectrophotometer drug content and encapsulation rate were measured. The characteristics of drug release in vitro which could simulate the actual state in vivo were tested by HPLC. Three ways of vein drop, artery perfusion and artery embolization were contrasted. Under the supervision of X-ray, PYM-GMS were perfused into the external carotid artery of rabbits by superselective artery embolization. Blood samples were tested at different time and analyzed statistically. Results: The roundness of PYM-GMS was 1.02?.005. The mean diameter was 85.6 mm, 78% of them ranging from 50-200 mm, which fitted the use of embolization. PYM content and encapsulation rate were 6.8% and 91.3% respectively. 70% of the drug was released in 3 h in the simulated environment in vivo and total drug was released after more than 6 h. After artery embolization with small dosage of PYM-GMS, the local drug concentration was 8 times higher than the blood drug concentration and the high level of local drug concentration was kept for more than 120 min. Conclusion: External carotid artery embolization with PYM-GMS, which significantly reduced the circulating drug level and employment dosage, could prolong the duration higher drug concentration and suit the purpose of targeted tumor therapy.

Preparation of Lung Targeting Microspheres of Gelatin Ceftiofur Alkali

Gelatin ceftiofur alkali microsphere was prepared to observe its characteristics and evaluate preservation conditions. The glutaraldehyde was increased and the carboxylic methyl chitosan was added to improve the microsphere. The experimental results show microspheres have a better morphology surface and fairly regular structure with 4% glutaraldehyde. The average particle size is 15.84 gm and particle size distribution is narrow which shows a good uniformity. Microsphere size was affected by the stirrer speed, dosing ratio and curing degree. The greater drug loaded is, the better microspheres loading is; but with the increase of drug loading rate, the entrapment efficiency increases first and then decreases. The drug release rate of the microsphere is 24.90% in 0.5 h and 84.90% in 48 h, when CMC-GMs with 4% curing agent is 32.03% in 0.5 h and 88.44% in 48 h. So Gms embedding of ceftiofur alkali are better than CMC-GM. The stability tests show that strong light, high temperature, high humidity have a great influence on the microspheres.

Preparation of magnetic gelatin-starch microspheres and adsorption performance for bovine serum album

The magnetic gelatin-starch microspheres were prepared by modified emulsion cross-linking method with glutaraldehyde as the cross-linking agent. The structure, size distribution as well as morphology of magnetic microspheres were investigated by FT-IR spectrometer, dynamic laser scattering analyzer and scanning electron microscope, respectively. Bovine serum album(BSA)was chosen as model protein, and the adsorption processes were carried out under diversified conditions including BSA initial concentration, p H value, adsorption time and temperature to evaluate the performance of the magnetic microspheres. The average diameter of optimized spherical magnetic microspheres is 1.6 μm with excellent dispersivity, and the saturation magnetization is found to be equal to 1.056×10-2 A·m2. The adsorption isotherm of the BSA on the magnetic microspheres basically obeys the Langmuir model, with a maximum adsorption capacity of 120 mg/g and an adsorption equilibrium constant of 1.60 mL/mg.

EXPERIMENTAL STUDY OF INTRATUMOR INJECTION WITH GELATIN MICROSPHERES CONTAINING ^(131)I AND MITOMYCIN C IN IMPLANTED HEPATOMA-22 IN MICE

The experiment of intratumor injection with gelatin microsphere containing 131I and mitomycin C (131I-MMC-GM) into implanted hepatoma-22 In mice is reported. Seventy Bal B/C mice were grouped into A, B, C, and D. Intratumor injection were given as follows: (A) 131I-MMC-GM, (B) 131I solution; (C) mitomycin C solution; (D) untreated control. The tumor-regression rates of the Group A, B and C as compared to Group D were 58. 7% , 23. 9% and 25. 4%. The average life times of Group A, B, C and were 40. 5, 25. 5, 24. 5 and 17. 1 days. Radioactivity counts in tumors and other organs in group A and B were measured with Y-Counter, which showed that 131I was concentrated in tumors in Group A but it was very low in other organs. The study showed that 131I-MMC-GM is effective and safe anticancer agent, intratumor injection with 131I-MMC-GM will be a promising therapy for the treatment of hepatoma.

Optimization of the Process of Gelatin-ceftiofur Sodium Microspheres

Gelatin microsphere（GMS） was prepared through W/O emulsion chemical-crossline method.The best formula was selected by examining its appearance,size,drug carrier and drug dissolution rate.The experimental results showed that the optimized gelatin microspheres were spherical ball with smooth surface and had well dispersion.The average size of blank gelatin microspheres was 15.84 μm,while the loaded microspheres＇average diameter were 33.10 μm.It was also shown that drug loading of microspheres increased with increasing loading capacity,but drug encapsulation efficiency had a trend of climbing up and then decline.The encapsulation efficiency reached the maximum when the dosage ratio was 2：1.And the results show ceftiofur sodium microspheres have sustained release in the PBS buffer of pH7.4.

Comparison of ethanol-soaked gelatin sponge and microspheres for hepatic arterioportal fistulas embolization in hepatic cellular carcinoma

Hepatic arterioportal fistulas(APFs)are common in hepatocellular carcinoma(HCC).Moreover,correlated with poor prognosis,APFs often complicate antitumor treatments,including transarterial chemoembolization(TACE).AIM To compare the efficacy of ethanol-soaked gelatin sponges(ESG)and microspheres in the management of APFs and their impact on the prognosis of HCC.METHODS Data from patients diagnosed with HCC or hepatic APFs between June 2016 and December 2019 were retrospectively analyzed.Furthermore,APFs were embolized with ESG(group E)or microspheres(group M)during TACE.The primary outcomes were disease control rate(DCR)and objective response rate(ORR).The secondary outcomes included immediate and first follow-up APF improvement,overall survival(OS),and progression-free survival(PFS).RESULTS Altogether,91 participants were enrolled in the study,comprising 46 in group E and 45 in group M.The DCR was 93.5%and 91.1%in groups E and M,respectively(P=0.714).The ORRs were 91.3%and 66.7%in groups E and M,respectively(P=0.004).The APFs improved immediately after the procedure in 43(93.5%)patients in group E and 40(88.9%)patients in group M(P=0.485).After 2 mo,APF improvement was achieved in 37(80.4%)and 33(73.3%)participants in groups E and M,respectively(P=0.421).The OS was 26.2±1.4 and 20.6±1.1 mo in groups E and M,respectively(P=0.004),whereas the PFS was 16.6±1.0 and 13.8±0.7 mo in groups E and M,respectively(P=0.012).CONCLUSION Compared with microspheres,ESG embolization demonstrated a higher ORR and longer OS and PFS in patients of HCC with hepatic APFs.

Effect of porous titanium coated with IGF-1 and TGF-β_1 loaded gelatin microsphere on function of MG63 cells

Porous titanium with porosity of 60% was prepared by metal injection molding（MIM）,and coated with gelatin sustained-release microspheres which were made by improved emulsified cold condensation method.The effects of porous titanium coated with insulin-like growth factor-1（IGF-1） and transforming growth factor-β1（TGF-β1） gelatin microspheres on the function of MG63 cells were evaluated in vitro.The results show that porous titanium coated with gelatin sustained-release microspheres has no cytotoxicity.The IGF-1 and TGF-β1 loading concentrations are positively correlative with the proliferation and differentiation of MG63 after co-culturing with the concentrations of IGF-1 and TGF-β1 gelatin microspheres in the range of 0.1-10 ng/mg and 0.25-2.5 ng/mg,respectively.The MG63 cells exhibit the best proliferation and differentiation with the IGF-1 and TGF-β1 loading concentrations of 10 ng/mg and 2.5 ng/mg,respectively.The joint application of IGF-1 and TGF-β1 group,which promote adhesion,proliferation and differentiation of MG63 cells,is superior to a single application group.

甲基丙烯化明胶微球缓释Kartogenin修复髓核退变的体外评估

背景:细胞外基质合成与降解的失衡是髓核退变的主要原因,小分子药物Kartogenin(KGN)可恢复基质合成和降解的平衡。利用合适的载药系统实现KGN的缓释对KGN发挥长期有效的治疗至关重要。目的:用甲基丙烯酰化明胶(gelatin methacryloyl,GelMA)包裹KGN,通过微流控技术制备成可注射的水凝胶微球,探究其生物相容性和对髓核细胞生物学功能的影响。方法:将β-环糊精(β-cyclodextrins,β-CD)与KGN混合成包合物,与10%GelMA按1∶9的体积混合,利用微流控技术制备可注射水凝胶微球GelMA@β-CD@KGN,扫描电镜表征微球的微观形貌,检测水凝胶微球浸泡于PBS中1个月的药物释放情况。提取SD大鼠髓核细胞,将第1代细胞分3组培养:对照组单独培养髓核细胞,另外两组分别将GelMA@β-CD微球、GelMA@β-CD@KGN微球与髓核细胞共培养,利用CCK-8法检测细胞增殖,活死细胞染色检测细胞存活。分别用含有白细胞介素1β和不含白细胞介素1β的完全培养基将髓核细胞与两种微球共培养,采用RT-PCR法检测髓核细胞基质合成蛋白和分解蛋白的mRNA表达。结果与结论:①扫描电镜下可见,冻干后的GelMA@β-CD@KGN微球为规则的球形,保持高度分散且大小均一,形状饱满;GelMA@β-CD@KGN微球体外可持续释放药物,至30 d时药物释放量达到总量的62%;②活死细胞染色显示,GelMA@β-CD@KGN可保持髓核细胞的活性;CCK-8检测显示,GelMA@β-CD@KGN可促进髓核细胞的增殖;③在含或不含白细胞介素1β的完全培养基中,GelMA@β-CD@KGN微球组聚集蛋白聚糖、Ⅱ型胶原的mRNA表达均高于GelMA@β-CD微球组(P<0.05,P<0.01),基质金属蛋白酶13、血小板反应蛋白解整合素金属肽酶5的mRNA表达均低于GelMA@β-CD微球组(P<0.01);④结果表明,GelMA@β-CD@KGN微球具有良好的生物相容性和药物缓释能力,作为载药系统是一种具有广阔应用前景的生物材料。

载槲皮素明胶微球对MC3T3-E1增殖和分化的影响

目的研究负载槲皮素的明胶三维多孔(G-quercetin)微球作为骨组织材料的可行性。方法利用乳化法制备负载槲皮素的多孔明胶微球,扫描电镜观察微球形态,通过免疫荧光染色、活死细胞染色和CCK-8、碱性磷酸酶(ALP)染色及茜素红染色检测微球的细胞毒性及其对小鼠胚胎成骨细胞前体细胞(MC3T3-E1)黏附、增殖及分化的影响,RT-PCR检测成骨相关基因Runx-2、ALP、OPN、OCN表达。结果扫描电镜结果显示制备的载槲皮素明胶三维微孔材料具有多孔结构。细胞黏附检测显示细胞能够在微球表面铺展良好。与对照组相比,活死细胞染色、CCK-8结果显示该微球无明显细胞毒性(P>0.05);与G-quercetin微球共培养的MC3T3-E1 ALP表达和体外矿化增加;PCR结果显示Runx-2、ALP、OCN、OPN表达明显增高(P<0.05)。结论载槲皮素明胶微球具有良好的生物相容性,能够促进MC3T3-E1体外成骨分化,有望作为新型骨组织工程生物材料应用于临床。

Preparation and in vitro evaluation of gelatin microspheres for embolization

The objective of this research was to develop gelatin microspheres（GMSs） and study their properties for embolization.The GMSs were prepared by emulsion chemical crosslinking method.The morphology and particle size of GMSs were observed under an optical microscope.The ratio of water absorption and GMSs swelling ratio were measured.The elasticity of GMSs was studied by TMS-Pro food texture analyzer.The deliverability of GMSs through catheter was investigated with a new device designed in this study.Finally,the acetone residue was determined by headspace capillary gas chromatography.The dried GMSs were elliptic with corrugated surface and the wet GMSs were round with smooth surface.The average diameter of dried GMSs was 430.5 μm with a range of 100-1000 μm,and that of wet GMSs was 601.2 μm with a range of 150-1425 μm.The maximum ratio of water absorption was 590.7% and the average swelling ratio was 103.0%.The elasticity of GMSs was proven to be appropriate for embolization.Three subgroups of wet GMSs（100-450 μm,450-700 μm and 700-940 μm） were delivered through catheter with acceptable pressure.The content of residual acetone in GMSs was less than the minimum limit in the Chinese Pharmacopoeia 2010.Therefore,we concluded that the GMSs were suitable for embolization according to the applicable properties in vitro.And the methods for assessing the properties of GMSs in this study were going to be useful in the evaluation of embolic microspheres.

壳聚糖/明胶复合微球的制备及吸附性能研究

以壳聚糖和明胶为原料,span 80为乳化剂,戊二醛为交联剂,采用乳化交联法制备了壳聚糖/明胶复合微球并用于Cr(Ⅵ)的吸附。考察了复合微球的各制备因素对微球形貌的影响,通过单因素分析研究了壳聚糖与明胶的比例、乳化剂用量、乳化时间对复合微球吸附性能的影响。结果表明,壳聚糖与明胶的比例为1∶2、span 80用量为6mL、乳化时间为50min时,制备的复合微球对Cr(Ⅵ)的吸附性能良好,吸附量较大,Cr(Ⅵ)的去除率可达95.3%。

载FGF10的多孔明胶微球对脊髓损伤大鼠的神经修复作用机制研究

目的探讨载成纤维生长因子10(FGF10)的多孔明胶微球(GMSs)对脊髓损伤大鼠神经保护作用的机制。方法20只健康雄性SD大鼠(220~250g)随机分为假手术组、脊髓损伤组、FGF10组、FGF10-GMSs组,每组各5只。假手术组只咬除T_(9)~T_(10)棘突及椎板等骨性组织,不造成脊髓损伤。其余3组造成脊髓损伤后,用Hamilton微量注射器向FGF10组大鼠的损伤注入20μL FGF10,向FGF10-GMSs组大鼠的损伤处注射20μL FGF10负载的多孔明胶微球,脊髓损伤组和假手术组的大鼠注射等量的生理盐水。收集大鼠胸椎(T_(9)~T_(10))脊髓组织,分别采用离体释放试验评估FGF10-GMSs的持续释放。使用大鼠脊髓损伤评分(BBB评分)和斜坡测试评估FGF10-GMSs对SCI大鼠的运动功能改善效果。使用HE、Nissl染色、TUNEL试验及Western blot评估FGF10-GMSs的治疗效果。结果离体释放实验结果显示,普通明胶微球会在短时间内迅速释放FGF10,多孔明胶微球持续而缓慢地释放FGF10,未在初始48 h内观察到显著的爆发释放模式。BBB评分结果显示,与脊髓损伤组(5分)相比,第21天FGF10-GMSs组(12分)和FGF10组(9分)的大鼠均表现出良好的运动功能恢复(P<0.05),斜坡测试结果与BBB评分结果一致,FGF10组和FGF10-GMSs组均获得了明显的行为改变。HE、Nissl染色结果显示FGF10-GMSs组内的坏死、核崩解和浸润性多核白细胞数量较少,在脊髓腔内坏死组织的比例显著降低。Western blot测定结果显示FGF10-GMSs组中的细胞凋亡抑制程度比FGF10组中更加明显。TUNEL试验也产生类似的结果,相比FGF10组和脊髓损伤组,FGF10-GMSs组具有更加明显的抑制细胞凋亡的作用。结论FGF10-GMSs通过抑制神经细胞的凋亡,从而达到保护脊髓损伤大鼠神经细胞的作用,为脊髓损伤的治疗提供新的思路,对提高脊髓损伤的预后具有十分重要的意义。

单宁酸交联明胶微球的制备及载药性能研究

目的:以天然植物提取物单宁酸为交联剂,明胶为原料,采用乳化交联法制备单宁酸明胶微球,并对其制备工艺进行优化和评价。方法:以外观形貌、平均粒径为指标,通过单因素试验筛选结合正交设计试验优化单宁酸明胶微球的制备工艺,最终制备花青素装载的单宁酸明胶微球;采用pH值示差法测定明胶微球的药物包封率及载药量。结果:单宁酸交联的明胶载药微球成球性好、形态圆整,大小均匀,优化后的微球平均粒径为(74.83±1.16)μm,在酸性、碱性环境下具有一定的耐腐蚀性。同时载药明胶微球的包封率可达89.90%,载药量为3.68%。结论:经单宁酸交联的明胶微球形态好、粒径适宜,并具有较高的药物装载率,为单宁酸交联明胶微球的后续研究提供了实验基础。

姜黄素肺靶向明胶微球的制备及性质研究

目的:研究姜黄素肺靶向明胶微球的制备工艺,并考察其理化性质。方法:以可生物降解的明胶为载体,液体石蜡为油相,Span80为乳化剂,采用正交设计优化制备工艺,用乳化交联法制备姜黄素肺靶向明胶微球。结果:优选所制的姜黄素肺靶向明胶微球形态圆整,表面光滑,载药量为6.15%,包封率为75.5%,5～30μm占总数的86.6%。22h体外释放50%,48h释放达77%。结论:制备工艺稳定可行,所得姜黄素明胶微球具有良好的缓释效果。