Results of multiple ligament reconstruction after knee dislocation--A prospective study with 95 patients and minimum 2-year follow up.

BACKGROUND: There is still a lack of clinical data in arthroscopic treatment for treating multiple ligament injuries. This study aims to evaluate the clinical outcomes of patients with multiple ligament injuries undergoing treatment based on the classification stage and type of injury. METHODS: A prospective, clinical trial on multiple ligament injuries was planned, which included 95 patients (58 men and 37 women; age: 42.8 ± 11.9 [range, 18-63] years) from October 2017 to June 2018. Injuries were classified into three stages (emergency stage < 24 h; acute stage: 24 h to 3 weeks, and chronic stage: > 3 weeks) and six types (KD I-VI) based on injuries time and structures, which indicated appropriate treatments. The clinical outcomes were evaluated at 2, 4, 6, 8, and 12 weeks and at 6, 9, 12 months and 24 months after surgery. The final choices in efficacy index included International Knee Documentation Committee (IKDC) score, Lysholm score, visual analog scale (VAS) score, and range of motion. RESULTS: During the follow-up, all patients exhibited statistically significant functional improvement in the injured limb compared with their preoperative situation. The mean postoperative scores of acute stage patients at 2-year follow-up were IKDC subjective score, 77.54 ± 11.53; Lysholm score, 85.96 ± 9.39; Tegner score, 4.13 ± 1.08; and VAS score, 1.21 ± 0.76. The mean postoperative scores of chronic stage patents at 2-year follow-up were IKDC subjective score, 74.61 ± 12.38; Lysholm score, 81.71 ± 10.80; Tegner score, 3.96 ± 1.14; and VAS score, 1.71 ± 0.60. The IKDC subjective score, Lysholm score, and Tegner score were significantly improved (P < 0.01) and the VAS score was significantly decreased (P < 0.01) at 2-year follow-up. Regarding the multiple ligament injuries classification, patients with more structural damage in stages V and VI showed less progress in functional recovery than those in stages I-IV. CONCLUSIONS: This new classification with three stages and six types helps to identify the severity of injury and plan the management effectively. The outcomes were encouraging and the subjective functional results showed significant improvement at 2-year follow-up. STUDY DESIGN: Prospective clinical trial. LEVEL OF EVIDENCE: II.

Combination of granulocyte colony-stimulating factor and CXCR4 antagonist AMD3100 for effective harvest of endothelial progenitor cells from peripheral blood and in vitro formation of primitive endothelial networks.

Endothelial progenitor cells (EPC) derived from the circulation may be used to enhance neovascularization. Since the combination of granulocyte colony-stimulating factor (GCSF) and CXCR4 antagonist AMD3100 efficiently mobilizes hematopoietic stem cells into peripheral circulation, it may increase the pool of endogenously circulating EPC. We tested this hypothesis by administering GCSF and AMD3100 to adult rabbits and rats, isolating mononuclear cells from peripheral blood by Ficoll density gradient centrifugation, and characterizing the blood-derived EPC based on morphology, immunophenotyping, gene expression and other functional analyses. These EPC showed clonal growth similar to that of human umbilical vein endothelial cells when cultured in complete EGM-2 medium on collagen I-precoated culture plates. The EPC exhibited a typical cobblestone-like morphology and were relatively homogeneous by the third passage. The cells expressed the typical endothelial marker CD31 based on flow cytometry and fluorescence microscopy, formed capillary-like structures when cultured in Matrigel, internalized DiI-acetylated low-density lipoprotein, bound Ulex europaeus agglutinin-1, and expressed CD31 and several other endothelial markers (VEGFR2, VE-cadherin, Tie-2, eNOS, vWF) at significantly higher levels than bone marrow-derived mesenchymal stem cells. These results suggest that the combination of GCSF and AMD3100 can efficiently release stem cells into peripheral circulation and generate EPC that show the desired morphological, immunophenotypic and functional characteristics. This minimally invasive approach may be useful for autologous cell transplantation for postnatal neovasculogenesis and tissue repair.

[A Novel All-arthroscopic PCL Tibial Inlay Reconstruction].

OBJECTIVE: To design new apparatus and develop relative surgical procedures for all-arthroscopic tibial inlay PCL reconstruction. METHODS: The new apparatus consisting of three main components for total arthroscopic tibial inlay were designed and manufactured, and the relative surgical procedures were experimented on 6 cadaveric knees. RESULTS: The apparatus for tibial sloting including three components was designed and manufactured successfully, which could be used easily. The surgical procedures were proved to be feasible, which could be performed within a short time (mean, 78.5 min). The graft fixation was proved to be firm enough by mechanical tests. Image examination (X ray, CT and MRI) confirmed that the location of implants was appropriate. CONCLUSION: The newly designed apparatus and surgical procedures for all-arthroscopic tibial inlay seems feasible and useful.

[3D Reconstruction of PCL Based on MRI and CT Image Fusion].

OBJECTIVE: To reconstruct three-dimensional (3D) image of posterior cruciate ligament (PCL) based on MRI and CT image fusion. METHODS: CT and MRI scans were performed on 12 knees of young men. The Dicom data were extracted and unified. The outline of PCL on MRI imaging was drew and plugged into the CT data. Finally, the visible 3D image of PCL with adjacent bones was reconstructed. The imaging anatomical measurements were examined and compared with those in published literature. RESULTS: Two cases were excluded from this study because of data deviations. The 3D visible reconstruction of PCL was proved to be feasible on the other ten cases. CONCLUSION: Three-dimensional visible reconstruction of PCL based on CT and MRI image fusion is feasible, which can provide support for individualized treatment of PCL injuries. Further simplification with increased accuracy may be needed.

Comparison of Anterior Cruciate Ligament Reconstruction With Versus Without Anterolateral Augmentation: A Systematic Review and Meta-analysis of Randomized Controlled Trials.

Background: It is clear that the anterolateral ligament has an important role in rotational knee stability. However, whether patients undergoing anterior cruciate ligament (ACL) reconstruction (ACLR) can benefit from anterolateral augmentation (ALA) is still controversial. Purpose: To compare the effects of isolated ACLR versus ACLR combined with ALA (ACLR+ALA) on clinical outcomes and knee stability. Study Design: Systematic review; Level of evidence, 1. Methods: The methodology followed PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. A literature search of the PubMed, Embase, and Cochrane Library Central Register of Controlled Trials databases was undertaken to identify all randomized controlled trials (RCTs) comparing isolated ACLR with ACLR+ALA for the treatment of ACL injuries in the last 15 years. The Cochrane Collaboration risk-of-bias tool and the revised Jadad scale were utilized by 2 independent reviewers to determine the quality of RCTs. Relevant data were extracted and compared between procedures, and heterogeneity across the RCTs was assessed using the I 2 statistic. Results: The initial search yielded 849 articles. A total of 14 studies (1850 patients; 941 ACLR and 909 ACLR+ALA) satisfied the eligibility criteria for the meta-analysis. There were no significant differences between the procedures in terms of patient-reported outcomes (International Knee Documentation Committee score, Tegner score, Knee injury and Osteoarthritis Outcome Score) or return-to-sport rates. However, patients who underwent ACLR+ALA had better knee stability based on the pivot-shift test (risk ratio [RR], 1.06 [95% CI, 1.02 to 1.10]; P = .0008), Lachman test (RR, 1.03 [95% CI, 1.00 to 1.07]; P = .04), and side-to-side difference in anterior laxity (standardized mean difference, -0.55 [95% CI, -0.98 to -0.12]; P = .01) as well as a lower incidence of graft failure (RR, 0.30 [95% CI, 0.19 to 0.45]; P < .01) compared with patients who underwent isolated ACLR. Conclusion: ALA can be considered as a reinforcement of ACLR to improve anteroposterior and anterolateral rotational stability of the knee and reduce the risk of failure. The patient-reported outcomes of isolated ACLR were similar to those of ACLR+ALA, and both procedures provided improved knee function.

[Modified single-needle arthroscopic repair technique for the treatment of longitudinal meniscal tear].

OBJECTIVE: To compare therapeutic efficacy of modified single-needle arthroscopic repair technique and Fast-Fix technique in repairing longitudinal meniscus injuries. METHODS: From July 2016 to July 2017, patients with longitudinal meniscus injuries who underwent meniscal repair surgery were retrospectively analyzed. Ninety-one patients treated with modified single-needle technique and 77 patients were treated with Fast-Fix technique, the average age were (26.7±7.6) and (27.9±6.1) years old respectively, the average lengths of follow-up were (32.5±9.2) and (33.2±11.9) months, respectively. Operation cost, suture time, intraoperative failure rate and postoperative failure rate were used as clinical outcomes, MRI of knee joint was used as main diagnosis and evaluation basis; 2000 IKDC subjective score, Lysholm score and Tegner activity scale were compared between two groups preoperatively, 12 months after operation and at the latest follow-up. Intraoperative and postopertaive complications were observed. RESULTS: Compared with Fast-Fix group, patients in modified single-needle technique group had lower operation costs [(645.7±133.1 vs.(12 184.8±4 709.8), P<0.01)], and average time per suture was shorter[(4.5±2.1) vs.(5.9±2.7), P<0.001)];and there were no significant difference in intraoperative failure rate and postoperative failure rate between two groups. There were no statistical differences in subjective function score, Lysholm score, Tegner activity knee joint at 12 months after opertaion and the latest follow-up (P>0.05);These scores between two groups at the latest followup were significantly improved compared with those of before operation(P<0.000 1). CONCLUSION: Modified single-needle arthrscopicrepair technique could achieve the similar therapeutic efficacy as Fast-Fix technique, and it has advantageds of simple opertion and more economical. This study recommends clinical application of modified single-needle arthrscopic repair technique in treating meniscus injuries.

Tß4 suppresses lincRNA-p21-mediated hepatic apoptosis and fibrosis by inhibiting PI3K-AKT-NF-κB pathway.

Hepatic injury is one of the most challenging diseases in clinical medicine. Hepatic injury is accompanied by hepatocyte apoptosis and leads to hepatic fibrosis and cirrhosis, which may cause liver cancer and increased mortality. Therefore, it is essential to investigate the regulation mechanism and therapeutic strategies for hepatic injury. In the study, the effects of Thymosin ß4 (Tß4) on Long intergenic noncoding RNA-p21 (lincRNA-p21)-mediated liver injury were investigated. Results showed that lincRNA-p21 overexpression promoted hepatocytes apoptosis, which was blocked by Tß4. Besides, Tß4 reversed the levels of cleaved caspase-3 and caspase-9 induced by lincRNA-p21. LincRNA-p21 overexpression also caused the pathological injury and fibrosis in hepatic tissues and increased the levels of fibrosis-related proteins (Collagen I, α-SMA and TIMP-1), and induced hydroxyproline and ALT production. However, Tß4 reversed the effects of overexpression of lincRNA-p21 on hepatic injury and fibrosis. In vitro experiments, after lincRNA-p21 was overexpressed in hepatic stellate cells (HSCs), the proliferation ability and the levels of HSCs markers α-SMA and Desmin were increased. However, Tß4 reversed the effects of lincRNA-p21 on HSCs. Furthermore, the PI3K-AKT-NF-κB pathway was activated by lincRNA-p21, which was then reversed by the Tß4 administration. After the mice treated by insulin-like growth factor-1 (IGF-1) (the activator of PI3K-AKT), the inhibitory effect of Tß4 on activated the PI3K-AKT-NF-κB pathway was abrogated. Besides, IGF-1 abolished the protective effects of Tß4 on hepatic apoptosis and fibrosis induced by lincRNA-p21. Therefore, Tß4 reversed. lincRNA-p21-mediated liver injury through inhibiting PI3K-AKT-NF-κB pathway. Tß4 may be a promising drug for fibrosis therapy.

The Effect of Irreducible Dislocation on Functional Outcomes in Knees With Multiligament Injuries: A Matched-Cohort Analysis.

BACKGROUND: Most cases of bicruciate knee dislocation (KD) with associated posteromedial disruption (KD-IIIM) are reducible, but some cannot be reduced by closed reduction because of soft tissue incarceration. PURPOSE: To compare the clinical characteristics and functional outcomes of KD-IIIM injuries in patients with or without incarceration of soft tissue requiring open or arthroscopic reduction. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: This retrospective cohort study of patients with KD was conducted between January 2013 and December 2017 at a single large institution. We applied a 1:2 matching ratio between patients with irreducible KD-IIIM injuries (irreducible group; n = 14) and those with reducible KD-IIIM injuries (control group; n = 28). There were 13 patients in the irreducible group and 25 in the control group who completed follow-up (≥2 years) and were included in our analysis. The efficacy of treatment in patients with KD was evaluated based on range of motion, the Tegner score, the Lysholm score, and the International Knee Documentation Committee (IKDC) score. RESULTS: At the end of follow-up, the mean Tegner score was 4.5 (range, 4-6), the mean Lysholm score was 79.2 (range, 60-95), and the mean IKDC score was 78.6 (range, 60.9-95.4) in the irreducible group. The respective results in the control group were 4.6 (range, 3-8), 83.1 (range, 39-100), and 80.6 (range, 42.5-96.6). These scores did not differ significantly between the 2 groups. Similarly, mean range of motion was similar between groups (irreducible, 118.1°; control, 124.8°). In the irreducible group, the acute subgroup showed significantly higher Lysholm and IKDC scores than the chronic subgroup, while the acute and chronic subgroups in the control group showed no significant differences in these respective outcome scores. CONCLUSION: In the present study, the treatment of irreducible KD led to similar functional outcomes compared with reducible KD. However, the treatment of chronic irreducible KD led to worse outcomes compared with acute irreducible KD, and therefore, urgent reduction is recommended in these patients.

Endocrine Therapy for the Functional Recovery of Spinal Cord Injury.

Spinal cord injury (SCI) is a major cause of physical disability and leads to patient dissatisfaction with their quality of life. Patients with SCI usually exhibit severe clinical symptoms, including sensory and motor dysfunction below the injured levels, paraplegia, quadriplegia and urinary retention, which can exacerbate the substantial medical and social burdens. The major pathological change observed in SCI is inflammatory reaction, which induces demyelination, axonal degeneration, and the apoptosis and necrosis of neurons. Traditional medical treatments are mainly focused on the recovery of motor function and prevention of complications. To date, numerous studies have been conducted to explore the cellular and molecular mechanism of SCI and have proposed lots of effective treatments, but the clinical applications are still limited due to the complex pathogenesis and poor prognosis after SCI. Endocrine hormones are kinds of molecules that are synthesized by specialized endocrine organs and can participate in the regulation of multiple physiological activities, and their protective effects on several disorders have been widely discussed. In addition, many studies have identified that endocrine hormones can promote nerve regeneration and functional recovery in individuals with central nervous system diseases. Therefore, studies investigating the clinical applications of endocrine hormones as treatments for SCI are necessary. In this review, we described the neuroprotective roles of several endocrine hormones in SCI; endocrine hormone administration reduces cell death and promotes functional repair after SCI. We also proposed novel therapies for SCI.

Evaluation of Decellularized Bovine Tendon Sheets for Achilles Tendon Defect Reconstruction in a Rabbit Model.

BACKGROUND: Achilles tendon (AT) defects frequently occur in trauma and chronic injuries. Currently, no method can satisfactorily reconstruct the AT with completely restored function. PURPOSE: To evaluate the postoperative outcomes of AT defect reconstruction with decellularized bovine tendon sheets (DBTSs) in a rabbit model. STUDY DESIGN: Controlled laboratory study. METHODS: DBTSs were prepared from bovine tendons after compression, decellularization, antigen extraction, freeze drying, and sterilization. Platelet-rich plasma (PRP) was obtained by differential centrifugation. Sixty-three rabbits were used in this study, and the AT defect model was created bilaterally. All rabbits were divided into 3 groups (n = 21). In the DBTS group and the DBTS + PRP group, 2-cm-long AT was excised and reconstructed by DBTSs or PRP-treated DBTSs. In the control group, the rabbits underwent AT transection, and stumps were sutured. After surgery, all rabbits were assessed by ultrasonography and magnetic resonance imaging and then sacrificed for histological examination and biomechanical testing at 4, 8, or 12 weeks. RESULTS: Gross observations demonstrated the absence of immunologic incompatibility and rejection. Histological examination showed that DBTSs promoted host cell infiltration and new fibrous tissue integration as compared with the control group. In each group, there was an AT-like structure formation and aligned collagen fiber deposition at 12 weeks. Mechanical properties of the reconstructed AT were not significantly different among the 3 groups at 4, 8, and 12 weeks after surgery ( P > .05). Ultrasonography and magnetic resonance imaging results illustrated that the reconstructed AT from each group maintained remodeling, and there was no significant difference in the echogenicity scoring ( P > .05) and percentages of good and excellent ( P > .05) among the 3 groups. CONCLUSION: DBTSs, which retain the native tendon structure and bioactive factors, had the ability to remodel and integrate into the rabbit AT and improve the healing process. CLINICAL RELEVANCE: DBTSs could serve as an effective bioscaffold to reconstruct AT defects.

[The prevention of hepatolithiasis and biliary stricture post choledochojejunostomy].

OBJECTIVE: To investigate the prevention of hepatolithiasis and biliary stricture post choledochojejunostomy using choledochoscopy technique and evaluate feasibility and efficacy of choledochojejunostomy and artificial valve of efferent loop in preventing reflux. METHODS: To analyze the data of 47 patients with hepatolithiasis who had been operated with Roux-en-Y cholangiojejunostomy and artificial valve of efferent loop to prevent bilio-intestinal regurgitation. Of the patients, 19 were marked with silver nip at the jejunum export of bilio-intestinal anastomosis. The regurgitation, recurrence, anastomotic stricture and their managements after the surgery were investigated. RESULTS: The bilio-intestinal regurgitation were found in 32 cases (32/47, 68.1%), it suggested that artificial valve could not prevent bilio-intestinal regurgitation efficiently. Two cases of hepatolithiasis recurred and were cured by sinus tract placement with the aid of silver nip mark under choledochoscope instead of re-operation. Of the 6 cases with anastomotic stricture, 5 cases were treated successfully with stone extraction, biliary stent dilatation under the percutaneous transhepatic cholangioscopy (PTCS) and the other one case died. CONCLUSIONS: Silver nip mark provides safe and simple path for the cholangioscopy, it made the treatment of the recurrent biliary stricture and stone safe and brief, made the cholangioscopy play more important role in the hepatolithiasis, residual stone and biliary stricture. PTCS is mini-invasive, safe, simple and effective.

[Clinical epidemiological analysis on posterior cruciate ligament injuries].

OBJECTIVE: To investigate clinical characteristics of posterior cruciate ligament (PCL) injuries. METHODS: To review all 326 patients with PCL injuries from January 2006 to June 2013 retrospectively, which were grouped according to injury patterns. The survey included the gender, age, sides, duration from injury to surgery, cause of injury and combined injuries. Compare the patient-specific factors between two groups. RESULTS: Male patients were the majority of PCL injuries (73%), and 30 to 50 years old patients accounted for 59%. 62.6% of all patients went for a doctor within one month after trauma. Traffic accidents were the main cause of PCL injuries (59%), especially motorcycle accidents (74.3%). Anterior cruciate ligament (ACL) injuries were the most common combined injuries (46.9%),followed by medial collateral ligament (MCL) (29.8%) and posterolateral corner (PLC) (26.1%). The meniscal tears accounted for 30.4% of total cases. The in-substance PCL injuries (71.5%) were far more than avulsion fractures (28.5%), and the former had more combined injuries than the later (P < 0.05). The causes of injuries were statistically different between two groups (P < 0.05). The prevalence of isolated avulsions was higher than isolated in-substance injuries (P < 0.05). CONCLUSION: In the general population, in-substance PCL tears were dominant on PCL injuries. The prevalence of combined injuries was quite high, among which ACL injuries were the most common. Insertion avulsion fractures emerged mainly at the tibial side, which had less combined injuries than those suffered in-substance PCL injuries. The incidence of isolated insertion avulsion fractures was higher than isolated in-substance injuries.

Mesenchymal Stem Cells Derived from Peripheral Blood Retain Their Pluripotency, but Undergo Senescence During Long-Term Culture.

Peripheral blood-derived mesenchymal stem cells (PB-MSCs) show promise as a source of cells for autologous transplantation because they can be harvested through minimally invasive procedures. To ensure adequate numbers of cells for transplantation and tissue regeneration, PB-MSCs must first be cultured and expanded in vitro, but whether long-term passage modifies their properties has been poorly understood. In this study we triggered production of PB-MSCs in rabbits using granulocyte colony-stimulating factor (G-CSF) and AMD3100, and then isolated and expanded the cells in culture until they reached a state of senescence, usually after about 20 passages. Cultures of low-, middle-, and high-passage numbers were compared in terms of morphology, proliferative capacity, phenotype, differentiation potential, apoptosis, metabolic indicators, and senescence. As passage number increased, MSCs retained their elongated spindle shape, but became larger and flatter, slowed in growth gradually, and increased proportion of cells showed G1 arrest. The proportions of apoptotic cells, production of reactive oxygen species (ROS), and ADP/ATP ratio increased with passage number. Expression of senescence-associated ß-galactosidase increased, while telomerase activity decreased. On the other hand, cultures did not show significant changes in phenotype or lose their ability to differentiate into three lineages as passage number increased. These results suggest that PB-MSCs maintain their stem cell properties during prolonged culturing, but they undergo senescence that may be due to apoptosis and production of ROS. These findings may help to standardize in vitro production of PB-MSCs for tissue engineering.

Coculture of peripheral blood-derived mesenchymal stem cells and endothelial progenitor cells on strontium-doped calcium polyphosphate scaffolds to generate vascularized engineered bone.

Vascularization of engineered bone tissue is critical for ensuring its survival after implantation and it is the primary factor limiting its clinical use. A promising approach is to prevascularize bone grafts in vitro using endothelial progenitor cells (EPC) derived from peripheral blood. Typically, EPC are added together with mesenchymal stem cells (MSC) that differentiate into osteoblasts. One problem with this approach is how to promote traditional tissue engineering bone survival with a minimally invasive method. In this study, we examined the effectiveness of administering to stimulate the release of peripheral blood stem cells and their co-culturing system for generating prevascularized engineered bone. Cells were isolated by Ficoll density gradient centrifugation and identified as EPC and MSC based on morphology, surface markers, and functional analysis. EPC and MSC were cocultured in several different ratios, and cell morphology and tube formation were assessed by microscopy. Expression of osteogenesis and vascularization markers was quantified by enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction, and histochemical and immunofluorescence staining. Increasing the proportion of EPC in the coculture system led to greater tube formation and greater expression of the endothelial cell marker CD31. An EPC:MSC ratio of 75:25 gave the highest expression of osteogenesis and angiogenesis markers. Cocultures adhered to a three-dimensional scaffold of strontium-doped calcium polyphosphate and proliferated well. Our findings show that coculturing peripheral blood-derived EPC and MSC may prove useful for generating prevascularized bone tissue for clinical use.

A new source of mesenchymal stem cells for articular cartilage repair: MSCs derived from mobilized peripheral blood share similar biological characteristics in vitro and chondrogenesis in vivo as MSCs from bone marrow in a rabbit model.

BACKGROUND: Bone marrow (BM) has been considered as a major source of mesenchymal stem cells (MSCs), but it has many disadvantages in clinical application. However, MSCs from peripheral blood (PB) could be obtained by a less invasive method and be more beneficial for autologous transplantation than BM MSCs, which makes PB a promising source for articular cartilage repair in clinical use. PURPOSE: To assess whether MSCs from mobilized PB of New Zealand White rabbits have similar biological characteristics in vitro and chondrogenesis in vivo as BM MSCs. STUDY DESIGN: Controlled laboratory study. METHODS: A combined method of drug administration containing granulocyte colony stimulating factor (G-CSF) plus CXCR4 antagonist AMD3100 was adopted to mobilize the PB stem cells of adult New Zealand White rabbits in vitro. The isolated cells were identified as MSCs by morphological characteristics, surface markers, and differentiation potentials. A comparison between PB MSCs and BM MSCs was made in terms of biological characteristics in vitro and chondrogenesis in vivo. This issue was investigated from the aspects of morphology, immune phenotype, multiple differentiation capacity, expansion potential, antiapoptotic capacity, and ability to repair cartilage defects in vivo of PB MSCs compared with BM MSCs. RESULTS: Peripheral blood MSCs were successfully mobilized by the method of combined drug administration, then isolated, expanded, and identified in vitro. No significant difference was found concerning the morphology, immune phenotype, and antiapoptotic capacity between PB MSCs and BM MSCs. Significantly, MSCs from both sources compounded with decalcified bone matrix showed the same ability to repair cartilage defects in vivo. For multipluripotency, BM MSCs exhibited a more osteogenic potential and higher proliferation capacity than PB MSCs, whereas PB MSCs possessed a stronger adipogenic and chondrogenic differentiation potential than BM MSCs in vitro. CONCLUSION: Although there are some differences in the proliferation and differentiation aspects between the 2 sources, PB MSCs share certain similar biological characteristics in vitro and chondrogenesis in vivo as BM MSCs. CLINICAL RELEVANCE: These results suggest that PB MSCs are a new source of seed cells used in articular cartilage repair.

Repair of large full-thickness cartilage defect by activating endogenous peripheral blood stem cells and autologous periosteum flap transplantation combined with patellofemoral realignment.

Minimal-invasive procedure and one-step surgery offer autologous mesenchymal stem cells derived from peripheral blood (PB-MSCs) a promising prospective in the field of cartilage regeneration. We report a case of a 19-year-old male athlete of kickboxing with ICRS grade IV chondral lesions at the 60° region of lateral femoral trochlea, which was repaired by activating endogenous PB-MSCs plus autologous periosteum flap transplantation combined with correcting the patellofemoral malalignment. After a 7.5 year follow-up, the result showed that the patient returned to competitive kickboxing. Second-look under arthroscopy showed a smooth surface at 8 months postoperation. The IKDC 2000 subjective score, Lysholm score and Tegner score were 95, 98 and 9 respectively at the final follow up. CT and MRI evaluations showed a significant improvement compared with those of pre-operation.

Comparative study of the biological characteristics of mesenchymal stem cells from bone marrow and peripheral blood of rats.

Mesenchymal stem cells (MSCs) from adult exhibit self-renewal and multilineage differentiation capacities, making the MSCs promising candidates for cell therapy and tissue engineering. Although bone marrow (BM) is the most universal source of MSCs, other tissues may also contain MSCs. Peripheral blood (PB), in particular, arises as the most attractive source of MSCs due to easy accessibility and noninvasive procedure. However, it is not certain that PB-MSCs have the equal biological characteristics to those of BM-MSCs. The purpose of this study was to compare the biological characteristics between BM-MSCs and PB-MSCs. We adopted granulocyte colony-stimulating factor combined with CXCR4 antagonist AMD3100 to stimulate MSCs to release into blood circulation of the rats. PB-MSCs were obtained from mobilized PB and expanded in long-term culture. BM-MSCs were isolated from the femur and tibia medullary canal of the same rats by density gradient centrifugation. After cell expansion in vitro, cell surface markers and multipotentiality analysis were performed to identify MSCs. Apoptosis resistance to H(2)O(2)-induced apoptosis, proliferation kinetics, cellular senescence, and karyotype analysis were measured to compare the biological characteristics of PB-MSCs and BM-MSCs. PB-MSCs with the typical adherent fibroblast-like morphology were similar to that of BM-MSCs. Both PB-MSCs and BM-MSCs were positive for CD44 and CD90, and negative for CD34 and CD45. They both exhibited trilineage differentiation potential and expressed lineage-specific genes. Although the BM-MSCs showed stronger osteogenic and adipogenic differentiation, PB-MSCs displayed a more chondrogenic capacity. Further, BM-MSCs have greater proliferation ability. Apoptosis resistance and cellular senescence were similar in MSCs derived from both sources. The results of our study demonstrate that PB-MSCs have similar biological characteristics to those of BM-MSCs despite certain minor differences, suggesting PB as a possible alternative source for MSCs.

Proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood under the culture conditions of hypoxia and serum deprivation.

BACKGROUND: The proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood (PB-MSCs) were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the feasibility for autologous PB-MSCs applications in cartilage repair. METHODS: MSCs were mobilized into peripheral blood by granulocyte colony stimulating factor (G-CSF) and AMD3100. The blood samples were collected from central ear artery of rabbits. Adhered cells were obtained by erythrocyte lysis buffer and identified as MSCs by adherence to plastic, spindle shaped morphology, specific surface markers, differentiation abilities into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. MSCs were cultured in four groups at different oxygen tension (20% O2 and 2% O2), with or without 10% fetal bovine serum (FBS) conditions: 20% O2 and 10% FBS complete medium (normal medium, N), 20% O2 and serum deprivation medium (D), 2% O2 and 10% FBS complete medium (hypoxia, H), 2% O2 and serum deprivation (HD). Cell proliferation was determined by CCK-8 assay. Apoptosis was detected by Annexin V/PI and terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL) staining. RESULTS: Spindle-shaped adherent cells were effectively mobilized from peripheral blood by a combined administration of G-CSF plus AMD3100. These cells showed typical fibroblast-like phenotype similar to MSCs from bone marrow (BM-MSCs), and expressed a high level of typical MSCs markers CD29 and CD44, but lacked in the expression of hematopoietic markers CD45 and major histocompatibility complex Class II (MHC II). They could also differentiate into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. No significant morphological differences were found among the four groups. It was found that hypoxia could enhance proliferation of PB-MSCs regardless of serum concentration, but serum deprivation inhibited proliferation at the later stage of culture. Apart from that, hypoxia or serum deprivation could promote the apoptosis of PB-MSCs after 48 hours; the effect was stronger when these two conditions combined together. Furthermore, the effect of serum deprivation on apoptosis was stronger compared with that of hypoxia. CONCLUSIONS: PB-MSCs possess similar phenotypes as BM-MSCs. Their differentiation and proliferation abilities make them a new source of seed cells for ischemia-related cell therapy and tissue engineering in the field of the articular cartilage repair.