Currently, there is no effective strategy to promote functional recovery after a spinal cord injury. Collagen scaffolds can not only provide support and guidance for axonal regeneration, but can also serve as a bridge...Currently, there is no effective strategy to promote functional recovery after a spinal cord injury. Collagen scaffolds can not only provide support and guidance for axonal regeneration, but can also serve as a bridge for nerve regeneration at the injury site. They can additionally be used as carriers to retain mesenchymal stem cells at the injury site to enhance their effectiveness. Hence, we hypothesized that transplanting human umbilical cord-mesenchymal stem cells on collagen scaffolds would enhance healing following acute complete spinal cord injury. Here, we test this hypothesis through animal studies and a phase I clinical trial.(1) Animal experiments: Models of completely transected spinal cord injury were established in rats and canines by microsurgery. Mesenchymal stem cells derived from neonatal umbilical cord tissue were adsorbed onto collagen scaffolds and surgically implanted at the injury site in rats and canines;the animals were observed after 1 week–6 months. The transplantation resulted in increased motor scores, enhanced amplitude and shortened latency of the motor evoked potential, and reduced injury area as measured by magnetic resonance imaging.(2) Phase I clinical trial: Forty patients with acute complete cervical injuries were enrolled at the Characteristic Medical Center of Chinese People's Armed Police Force and divided into two groups. The treatment group(n = 20) received collagen scaffolds loaded with mesenchymal stem cells derived from neonatal umbilical cordtissues;the control group(n = 20) did not receive the stem-cell loaded collagen implant. All patients were followed for 12 months. In the treatment group, the American Spinal Injury Association scores and activities of daily life scores were increased, bowel and urinary functions were recovered, and residual urine volume was reduced compared with the pre-treatment baseline. Furthermore, magnetic resonance imaging showed that new nerve fiber connections were formed, and diffusion tensor imaging showed that electrophysiological activity was recovered after the treatment. No serious complication was observed during follow-up. In contrast, the neurological functions of the patients in the control group were not improved over the follow-up period. The above data preliminarily demonstrate that the transplantation of human umbilical cord-mesenchymal stem cells on a collagen scaffold can promote the recovery of neurological function after acute spinal cord injury. In the future, these results need to be confirmed in a multicenter, randomized controlled clinical trial with a larger sample size. The clinical trial was approved by the Ethics Committee of the Characteristic Medical Center of Chinese People's Armed Police Force on February 3, 2016(approval No. PJHEC-2016-A8). All animal experiments were approved by the Ethics Committee of the Characteristic Medical Center of Chinese People's Armed Police Force on May 20, 2015(approval No. PJHEC-2015-D5).展开更多
Background: Osteoarthritis is one of the most common bone diseases, triggered by bone destruction stemming from the inflammatory response of chondrocytes. The disease progresses slowly, but halting its progression or ...Background: Osteoarthritis is one of the most common bone diseases, triggered by bone destruction stemming from the inflammatory response of chondrocytes. The disease progresses slowly, but halting its progression or finding a cure remains elusive. The treatment of pain associated with osteoarthritis has yielded unsatisfactory results. In recent years, mesenchymal stem cells (MSCs) have emerged as a potential avenue for addressing the condition. In this study, we used MSCs to treat companion dogs with osteoarthritis. Methods: For this study, 26 animals were included in this study to assess the pain and mobility one month after treatment. The pain scores were obtained from owners using a questionnaire based on the Helsinki Chronic Pain Index, and the Liverpool Osteoarthritis in Dogs (LOAD) Owner questionnaire to assess the mobility of the dogs from stem cell infusion. Results: Questionnaires were administered to dog owners before and one month after treatment, and we found that dogs treated with MSCS experienced an 81.2% ± 6.8% reduction in pain and a 77.9% ± 10.1% increase in mobility, whereas most of the dogs in the untreated control group experienced disease progression. Conclusions: The transplantation of stem cells into companion pets is a promising and expanding opportunity for pet owners with aging and arthritic dogs. MSCs may play an important role in the treatment of OA without complications in companion pets.展开更多
Canine adipose derived stem cells (ASCs) hold a great promise for the therapy of osteoarthritis in veterinary medicine. Current therapy is an autologous, stromal vascular fraction. Allogeneic ASCs provide many advanta...Canine adipose derived stem cells (ASCs) hold a great promise for the therapy of osteoarthritis in veterinary medicine. Current therapy is an autologous, stromal vascular fraction. Allogeneic ASCs provide many advantages, including efficient, cost-effective treatments while eliminating a surgical procedure in a diseased animal. Cultured ASCs can be expanded and characterized, allowing selection of desirable qualities. Use of allogeneic ASCs requires selection of a culture medium that provides consistent, desirable cellular products. The supplements within a medium can greatly influence cellular phenotypes. We hypothesized that medium type influenced cellular phenotype, allowing selection of a specified cellular product for clinical applications. We evaluated ASCs derived from adipose tissue of six dogs, assessing mRNA expression of proinflammatory: interleukin-1b, cyclooxygenase-2, and anti-inflammatory mediators: tissue inhibitor metalloproteinase-2 and interleukin -1 receptor antagonist, via quantitative RT-PCR prior to, and following culture in five cell culture media: basic cell growth medium (BGM), Keratinocyte N acetyl-L-cysteine supplemented (KNAC) medium, Multipotent Adult Progenitor Cell (MAPC) medium, serum free medium (SFM) and xeno-free medium. Major histocompatability complex I (MHCI), major histocompatability complex II (MHCII), CD44 and CD90 immunophenotypes were assessed via flow cytometry analysis. Tri-lineage differentiation (bone, adipose and cartilage tissue) was utilized to verify multipotency. SFM and xeno-free culture conditions did not produce cell expansion sufficient to assess phenotype. ASCs prior to culture had wide variability in all mediator levels, while culturing in the remaining conditions resulted in more predictable expression levels of inflammatory mediators, with a decrease in all levels. Cultured ASCs retained expression of cell surface markers MHCI, CD44 and CD90, while decreasing MHCII expression levels. KNAC and MAPC medium conditions consistently produced tri-lineage differentation;BGM, SFM and xeno-free medium did not. Culture condition will influence phenotype of ASCs, and should be selected according to the intended therapeutic effect.展开更多
Recently,we read a mini-review published by Jeyaraman et al.The article explored the optimal methods for isolating mesenchymal stromal cells from adipose tissue-derived stromal vascular fraction(SVF).Key factors inclu...Recently,we read a mini-review published by Jeyaraman et al.The article explored the optimal methods for isolating mesenchymal stromal cells from adipose tissue-derived stromal vascular fraction(SVF).Key factors include tissue source,processing techniques,cell viability assessment,and the advantages/disadvantages of autologous vs allogeneic use.The authors emphasized the need for standardized protocols for SVF isolation,ethical and regulatory standards for cell-based therapy,and safety to advance mesenchymal stromal cell-based therapies in human patients.This manuscript shares our perspective on SVF isolation in canines.We discussed future directions to potentiate effective regenerative medicine therapeutics in human and veterinary medicine.展开更多
【目的】探究犬脐带间充质干细胞(UC-MSCs)联合滑车沟再造术对髌骨脱位治疗的效果。【方法】将20只髌骨脱位患犬随机分为干细胞治疗组和常规手术组,每组10只。手术当天,干细胞治疗组关节腔注射0.5 mL UC-MSCs(106/kg)混悬液,常规手术组...【目的】探究犬脐带间充质干细胞(UC-MSCs)联合滑车沟再造术对髌骨脱位治疗的效果。【方法】将20只髌骨脱位患犬随机分为干细胞治疗组和常规手术组,每组10只。手术当天,干细胞治疗组关节腔注射0.5 mL UC-MSCs(106/kg)混悬液,常规手术组注入等量生理盐水,通过收集患犬基础信息、回访跟踪记录,检测血常规和血液因子含量以及影像学等方法评估犬UC-MSCs的治疗效果。【结果】与常规手术组对比,术后第1、7天干细胞治疗组白细胞介素-6(IL-6)含量极显著或显著降低(P<0.01;P<0.05),两组间转化生长因子-β1(TGF-β1)、基质金属蛋白酶13(MMP-13)及肿瘤坏死因子α(TNF-α)和白细胞总数、中性粒细胞总数均无显著差异(P>0.05)。术后第30天数字X线摄影复查结果显示,干细胞治疗组关节腔清晰,骨损位置有明显的软骨及骨组织生长,无再次脱出和关节炎等并发症。常规手术组的骨损伤处骨生长情况缓慢,2例患犬髌骨再次脱位(2/10)。【结论】滑车沟再造术后,在常规术后护理(使用抗炎、抗菌和止疼药物)条件下,关节腔内注射UC-MSCs能促进软骨和骨组织生长,降低再次脱出发生的风险。UC-MSCs联合滑车沟再造术能提升犬髌骨脱位的临床治疗效果。展开更多
为了研究犬骨髓间充质干细胞(can ine m esenchym a l stem ce lls,cM SC s)的生长特点和在诱导条件下的成骨特性。使用密度梯度法分离成年犬骨髓间充质干细胞进行培养,保留贴壁细胞传代,观察,以地塞米松、β甘油磷酸钠、抗坏血酸为成...为了研究犬骨髓间充质干细胞(can ine m esenchym a l stem ce lls,cM SC s)的生长特点和在诱导条件下的成骨特性。使用密度梯度法分离成年犬骨髓间充质干细胞进行培养,保留贴壁细胞传代,观察,以地塞米松、β甘油磷酸钠、抗坏血酸为成骨诱导剂。利用倒置光学显微镜和透射电镜观察细胞形态特征,四甲基偶氮盐(M TT)比色测定增殖,用碱性磷酸酶(A lka line phosphatase,ALP)活性及骨钙素(O steoca lc in,OCN)含量来研究细胞分化情况。形态学观察表明,cM SC s贴壁细胞呈集落生长,有成纤维细胞样外观,透射电镜可见成骨诱导后cM SC s具有分泌型细胞的特征;推测成骨诱导剂可促进骨髓间充质干细胞成骨,表现为ALP活性、OCN含量明显升高。本实验表明所培养的cM SC s保持了未分化状态,并在成骨诱导剂的作用下可向成骨细胞分化。展开更多
基金supported by the National Natural Science Foundation of China,Nos.11932013(to SZ),11672332(to SZ)the National Key Research and Development Plan of China,No.2016YFC1101500(to SZ)+2 种基金the Science and Technology Military-Civilian Integration Project of Tianjin of China,No.18ZXJMTG00260(to XYC)the Key Project of Science and Technology Support Plan of Tianjin of China,No.17YFZCSY00620(to XYC)the Rescue Medical Clinical Center Fund of Tianjin of China,No.15ZXLCSY00040(to XYC)
文摘Currently, there is no effective strategy to promote functional recovery after a spinal cord injury. Collagen scaffolds can not only provide support and guidance for axonal regeneration, but can also serve as a bridge for nerve regeneration at the injury site. They can additionally be used as carriers to retain mesenchymal stem cells at the injury site to enhance their effectiveness. Hence, we hypothesized that transplanting human umbilical cord-mesenchymal stem cells on collagen scaffolds would enhance healing following acute complete spinal cord injury. Here, we test this hypothesis through animal studies and a phase I clinical trial.(1) Animal experiments: Models of completely transected spinal cord injury were established in rats and canines by microsurgery. Mesenchymal stem cells derived from neonatal umbilical cord tissue were adsorbed onto collagen scaffolds and surgically implanted at the injury site in rats and canines;the animals were observed after 1 week–6 months. The transplantation resulted in increased motor scores, enhanced amplitude and shortened latency of the motor evoked potential, and reduced injury area as measured by magnetic resonance imaging.(2) Phase I clinical trial: Forty patients with acute complete cervical injuries were enrolled at the Characteristic Medical Center of Chinese People's Armed Police Force and divided into two groups. The treatment group(n = 20) received collagen scaffolds loaded with mesenchymal stem cells derived from neonatal umbilical cordtissues;the control group(n = 20) did not receive the stem-cell loaded collagen implant. All patients were followed for 12 months. In the treatment group, the American Spinal Injury Association scores and activities of daily life scores were increased, bowel and urinary functions were recovered, and residual urine volume was reduced compared with the pre-treatment baseline. Furthermore, magnetic resonance imaging showed that new nerve fiber connections were formed, and diffusion tensor imaging showed that electrophysiological activity was recovered after the treatment. No serious complication was observed during follow-up. In contrast, the neurological functions of the patients in the control group were not improved over the follow-up period. The above data preliminarily demonstrate that the transplantation of human umbilical cord-mesenchymal stem cells on a collagen scaffold can promote the recovery of neurological function after acute spinal cord injury. In the future, these results need to be confirmed in a multicenter, randomized controlled clinical trial with a larger sample size. The clinical trial was approved by the Ethics Committee of the Characteristic Medical Center of Chinese People's Armed Police Force on February 3, 2016(approval No. PJHEC-2016-A8). All animal experiments were approved by the Ethics Committee of the Characteristic Medical Center of Chinese People's Armed Police Force on May 20, 2015(approval No. PJHEC-2015-D5).
文摘Background: Osteoarthritis is one of the most common bone diseases, triggered by bone destruction stemming from the inflammatory response of chondrocytes. The disease progresses slowly, but halting its progression or finding a cure remains elusive. The treatment of pain associated with osteoarthritis has yielded unsatisfactory results. In recent years, mesenchymal stem cells (MSCs) have emerged as a potential avenue for addressing the condition. In this study, we used MSCs to treat companion dogs with osteoarthritis. Methods: For this study, 26 animals were included in this study to assess the pain and mobility one month after treatment. The pain scores were obtained from owners using a questionnaire based on the Helsinki Chronic Pain Index, and the Liverpool Osteoarthritis in Dogs (LOAD) Owner questionnaire to assess the mobility of the dogs from stem cell infusion. Results: Questionnaires were administered to dog owners before and one month after treatment, and we found that dogs treated with MSCS experienced an 81.2% ± 6.8% reduction in pain and a 77.9% ± 10.1% increase in mobility, whereas most of the dogs in the untreated control group experienced disease progression. Conclusions: The transplantation of stem cells into companion pets is a promising and expanding opportunity for pet owners with aging and arthritic dogs. MSCs may play an important role in the treatment of OA without complications in companion pets.
文摘Canine adipose derived stem cells (ASCs) hold a great promise for the therapy of osteoarthritis in veterinary medicine. Current therapy is an autologous, stromal vascular fraction. Allogeneic ASCs provide many advantages, including efficient, cost-effective treatments while eliminating a surgical procedure in a diseased animal. Cultured ASCs can be expanded and characterized, allowing selection of desirable qualities. Use of allogeneic ASCs requires selection of a culture medium that provides consistent, desirable cellular products. The supplements within a medium can greatly influence cellular phenotypes. We hypothesized that medium type influenced cellular phenotype, allowing selection of a specified cellular product for clinical applications. We evaluated ASCs derived from adipose tissue of six dogs, assessing mRNA expression of proinflammatory: interleukin-1b, cyclooxygenase-2, and anti-inflammatory mediators: tissue inhibitor metalloproteinase-2 and interleukin -1 receptor antagonist, via quantitative RT-PCR prior to, and following culture in five cell culture media: basic cell growth medium (BGM), Keratinocyte N acetyl-L-cysteine supplemented (KNAC) medium, Multipotent Adult Progenitor Cell (MAPC) medium, serum free medium (SFM) and xeno-free medium. Major histocompatability complex I (MHCI), major histocompatability complex II (MHCII), CD44 and CD90 immunophenotypes were assessed via flow cytometry analysis. Tri-lineage differentiation (bone, adipose and cartilage tissue) was utilized to verify multipotency. SFM and xeno-free culture conditions did not produce cell expansion sufficient to assess phenotype. ASCs prior to culture had wide variability in all mediator levels, while culturing in the remaining conditions resulted in more predictable expression levels of inflammatory mediators, with a decrease in all levels. Cultured ASCs retained expression of cell surface markers MHCI, CD44 and CD90, while decreasing MHCII expression levels. KNAC and MAPC medium conditions consistently produced tri-lineage differentation;BGM, SFM and xeno-free medium did not. Culture condition will influence phenotype of ASCs, and should be selected according to the intended therapeutic effect.
基金Supported by the Department of Biotechnology,Ministry of Science and Technology,Government of India,New Delhi,No.BT/PR42179/AAQ/1/814/2021SERB-State University Research Excellence,No.SUR/2022/001952.
文摘Recently,we read a mini-review published by Jeyaraman et al.The article explored the optimal methods for isolating mesenchymal stromal cells from adipose tissue-derived stromal vascular fraction(SVF).Key factors include tissue source,processing techniques,cell viability assessment,and the advantages/disadvantages of autologous vs allogeneic use.The authors emphasized the need for standardized protocols for SVF isolation,ethical and regulatory standards for cell-based therapy,and safety to advance mesenchymal stromal cell-based therapies in human patients.This manuscript shares our perspective on SVF isolation in canines.We discussed future directions to potentiate effective regenerative medicine therapeutics in human and veterinary medicine.
文摘【目的】探究犬脐带间充质干细胞(UC-MSCs)联合滑车沟再造术对髌骨脱位治疗的效果。【方法】将20只髌骨脱位患犬随机分为干细胞治疗组和常规手术组,每组10只。手术当天,干细胞治疗组关节腔注射0.5 mL UC-MSCs(106/kg)混悬液,常规手术组注入等量生理盐水,通过收集患犬基础信息、回访跟踪记录,检测血常规和血液因子含量以及影像学等方法评估犬UC-MSCs的治疗效果。【结果】与常规手术组对比,术后第1、7天干细胞治疗组白细胞介素-6(IL-6)含量极显著或显著降低(P<0.01;P<0.05),两组间转化生长因子-β1(TGF-β1)、基质金属蛋白酶13(MMP-13)及肿瘤坏死因子α(TNF-α)和白细胞总数、中性粒细胞总数均无显著差异(P>0.05)。术后第30天数字X线摄影复查结果显示,干细胞治疗组关节腔清晰,骨损位置有明显的软骨及骨组织生长,无再次脱出和关节炎等并发症。常规手术组的骨损伤处骨生长情况缓慢,2例患犬髌骨再次脱位(2/10)。【结论】滑车沟再造术后,在常规术后护理(使用抗炎、抗菌和止疼药物)条件下,关节腔内注射UC-MSCs能促进软骨和骨组织生长,降低再次脱出发生的风险。UC-MSCs联合滑车沟再造术能提升犬髌骨脱位的临床治疗效果。
文摘为了研究犬骨髓间充质干细胞(can ine m esenchym a l stem ce lls,cM SC s)的生长特点和在诱导条件下的成骨特性。使用密度梯度法分离成年犬骨髓间充质干细胞进行培养,保留贴壁细胞传代,观察,以地塞米松、β甘油磷酸钠、抗坏血酸为成骨诱导剂。利用倒置光学显微镜和透射电镜观察细胞形态特征,四甲基偶氮盐(M TT)比色测定增殖,用碱性磷酸酶(A lka line phosphatase,ALP)活性及骨钙素(O steoca lc in,OCN)含量来研究细胞分化情况。形态学观察表明,cM SC s贴壁细胞呈集落生长,有成纤维细胞样外观,透射电镜可见成骨诱导后cM SC s具有分泌型细胞的特征;推测成骨诱导剂可促进骨髓间充质干细胞成骨,表现为ALP活性、OCN含量明显升高。本实验表明所培养的cM SC s保持了未分化状态,并在成骨诱导剂的作用下可向成骨细胞分化。
