Clinical application of individualized 3D-printed navigation template to children with cubitus varus deformity.

BACKGROUND: Cubitus varus deformity is a common sequela of elbow fractures in children. Cubitus varus deformity treatment is tending toward 3D correction, which is challenging for orthopedic surgeons. This study aims to explore whether individualized 3D-printed navigation templates can assist with accurate and effective corrective treatment of children with cubitus varus deformity. METHODS: Thirty-five patients were treated for cubitus varus deformity from June 2015 to April 2017, including 21 boys and 14 girls, aged 4.6-13.2 years (average, 7.5 years). Of these cases, 17 deformities were on the left side and 18 were on the right side. All were treated with wedge osteotomy of the lateral distal humerus. 3D-printed navigation templates were used in 16 cases, while traditional surgery was used in 19 cases. All patients underwent computed tomography scans before surgery. Computer software was used to analyze the measurements and design and print individualized navigation templates. The navigation templates were matched, and surgery was initially simulated. Intraoperative individualized navigation templates were used to assist with accurate osteotomy and Kirschner wire fixation. Operation times were recorded in all cases, the carrying angles before and after surgery were assessed by computer, and postoperative elbow joint function was evaluated using Bellemore criteria. All measurement data were presented as means ± SD, and Student's t test was used to examine differences between groups. All count data between both groups were compared using the chi-square test or Fisher's exact test analysis. RESULTS: All individualized navigation templates matched well with the corresponding anatomical markers and were consistent with preoperative planning, simulated surgery, and intraoperative procedures. Average operation times from clear exposure to fixed Kirschner wire were 11.69 min (9.6-13.5 min) for the individualized navigation template group and 22.89 min (17.7-26.8 min) for the traditional operation group (p < 0.001). Average differences in postoperation carrying angles between affected and healthy sides were 1.13° (0-2.0°) and 4.21° (0-7.5°), respectively (p < 0.001). Follow-up 6-12 months postoperation showed that elbow function did not differ significantly between groups using the Bellemore criteria (p > 0.05). CONCLUSIONS: Individualized navigation templates simplify procedures, reduce operation time, and improve accuracy when used in orthopedic surgery to treat children with cubitus varus deformity.

Application of 3D printing rapid prototyping-assisted percutaneous fixation in the treatment of intertrochanteric fracture.

The aim of the present study was to investigate the application of 3D printing (3DP) rapid prototyping (RP) technique-assisted percutaneous fixation in the treatment of femoral intertrochanteric fracture (ITF) using proximal femoral nail anti-rotation (PFNA). A total of 39 patients with unstable ITF were included in the current study. Patients were divided into two groups: 19 patients were examined using computed tomography scanning and underwent PFNA with SDP-RP whereas the other 20 patients underwent conventional PFNA treatment. Anatomical data were converted from the Digital Imaging and Communications in Medicine format to the stereolithography format using M3D software. The 3DP-RP model was established using the fused deposition modeling technique and the length and diameter of the main screw blade was measured during the simulation. The postoperative femoral neck-shaft angle (NSA), surgery duration, intraoperative and postoperative blood loss, and the duration of hospital stay were recorded and compared with the corresponding values in conventional surgery. No significant differences were observed in mean PFNA size between the implants used and the preoperative planning estimates. It was demonstrated that the 3DP-RP assisted procedure resulted in more effective reduction of the NSA. Furthermore, patients undergoing 3DP-RP experienced a significant reduction in duration of surgery (P<0.01), as well as reductions in intraoperative (P=0.02) and postoperative (P=0.03) blood loss, compared with conventional surgery. At 6 months post-surgery, no cases of hip varus/vague deformities or implant failure were observed in patients that underwent either the 3DP-RP-assisted or conventional procedure. The results of the present study suggest that the 3DP-RP technique is able to create an accurate model of the ITF, which facilitates surgical planning and fracture reduction, thus improving the efficiency of PFNA surgery for ITFs.

Adhesion, proliferation and osteogenic differentiation of mesenchymal stem cells in 3D printed poly-ε-caprolactone/hydroxyapatite scaffolds combined with bone marrow clots.

Mesenchymal stem cells (MSCs), a stem cell population capable of multi­lineage differentiation, bound to porous biomaterial scaffolds, are widely used for bone tissue regeneration. However, there is evidence to suggest that MSC collection from bone marrow and expansion in vitro may result in phenotypic changes including a loss of differentiation potential and cell senescence. The aim of the present study was to find a facile and efficient approach to enable MSC adhesion and proliferation to scaffolds with osteogenic differentiation. Unprocessed bone marrow blood from the condyle of the distal femur in the rabbits were added to three­dimensional (3D) printed porous poly-ε-caprolactone/hydroxyapatite (PCL/HA) scaffolds with bone marrow clots (MC) formed, using two different methods for Group A (MC enriched scaffolds) and Group B (MC combined scaffolds), and then were cultured in osteogenic medium for 4 weeks. The scaffolds were assessed macroscopically and microscopically. Scaffold bioactivity and the proliferation and osteogenic differentiation of seeded MSCs were measured. Higher cellular viability and greater cell numbers in the scaffolds at later phases of culture were observed in Group B compared with Group A. In addition, Group B was associated with greater osteoinductivity, alkaline phosphatase activity and bony nodule formation, as assessed using scanning electron microscopy. Furthermore, reverse transcription­quantitative polymerase chain reaction analysis revealed that more osteogenic differentiation was present in Group B, compared with Group A. MC combined scaffolds proved to be a highly efficient, reliable and simple novel method for MSC adhesion, proliferation and differentiation. The MC combined PCL­HA multi­scale porosity scaffold may represent a candidate for future bone regeneration studies.

Cartilage matrix changes in contralateral mobile knees in a rabbit model of osteoarthritis induced by immobilization.

BACKGROUND: Many researches have investigated the changes associated with immobilization-induced osteoarthritis (OA). However, there are only few studies focusing on the effect of unilateral knee immobilization on cartilage matrix changes in the contralateral mobile knee. The aim of the present study was to investigate the influence of immobilization on the cartilage matrix in the contralateral mobile knees in a rabbit model of OA induced by immobilization. METHODS: Right knees (experimental knees) of eighteen mature female rabbits were immobilized at an extension of 180° with orthopedic casting tape for 2, 4, or 8 weeks. Left knees (contralateral knees) of the immobilized rabbits were not subjected to immobilization. The knees of six non-immobilized rabbits were designated as control knees. Following immobilization, cartilage specimens from the medial femoral condyle underwent macroscopic, histological, immunohistochemical, and biochemical evaluations. RESULTS: Roughness of cartilage surface was detected in the experimental knees at 2 weeks, and cartilage degeneration was further developed. In the contralateral knee, cartilage showed degenerative changes after 4 weeks. Safranin-O staining and glycosaminoglycan (GAG) contents were reduced in the experimental knees following immobilization and in the contralateral intact knees after 4 and 8 weeks. Type II collagen staining was gradually reduced, type I collagen accumulation was obviously detected in the upper and middle layers of cartilage in experimental knees after 8 weeks, and the collagen orientation was gradually disorganized in both knees at 4 and 8 weeks. For both experimental and contralateral knees, collagen contents were significantly decreased at 8 weeks, and Mankin and Osteoarthritis Research Society International (OARSI) scores increased over time. CONCLUSION: OA developed in the contralateral intact knee with the progress of OA in the immobilized knee in a rabbit model of immobilization-induced OA.

Using 7.0T MRI T2 mapping to detect early changes of the cartilage matrix caused by immobilization in a rabbit model of immobilization-induced osteoarthritis.

OBJECTIVE: The goal of this study was to detect early changes in the cartilage matrix caused by immobilization in a rabbit model of immobilization-induced osteoarthritis (OA) by T2 mapping with 7.0T MRI. MATERIALS AND METHODS: Left knee joints of 28 mature rabbits were immobilized at 180° of extension with orthopedic casting tape for 1, 2, or 3weeks (n=7 rabbits each). No immobilization was performed in the control group (n=7 rabbits). T2 mapping was performed after 1, 2, and 3weeks. Osteochondral specimens harvested from the trochlea groove (TG) and medial femoral condyle (MFC) were subjected to histologic, immunohistochemical, and microscopic evaluation, followed by biochemical assays for water, glycosaminoglycan (GAG), and collagen. The ability of T2 mapping to reveal changes in the cartilage matrix was further assessed. RESULTS: Rabbits demonstrated elevated T2 values (9.9% in TG, 10.6% in MFC), a dulled cartilage surface, reduced Safranin-O staining, and decreased GAG content (14.2% in TG and MFC) after 2weeks, with cartilage surface softening, irregularity, and markedly reduced GAG content by 3weeks. T2 values were correlated positively with water (r=0.836 in TG, r=0.821 in MFC) and negatively with GAG content (r=-0.945 in TG, r=-0.957 in MFC), but had no discernible relationship with collagen content (r=-0.196 in TG, r=-0.213 in MFC). CONCLUSIONS: 7.0T MRI T2 mapping can be used to detect early changes of the cartilage matrix caused by immobilization in an immobilization-induced OA model.

Effects of nicotine on a rat model of early stage osteoarthritis.

The objective of the study was to investigate the effects of nicotine on articular cartilage degeneration and inflammation in a rat model of early stage osteoarthritis (OA), using T2 mapping. In this study, a rat model of early stage OA was established by immobilizing the left knee joints of adult male rats for two weeks. Subsequently, rats were fed with nicotine for two and four weeks. Changes in the articular cartilage from the medial femoral condylar region of the knee were evaluated by gross observation and histological grading with the contents of cartilage matrix detected. T2 values of the articular cartilage were estimated through high-field magnetic resonance imaging (MRI) (7.0 T). Levels of serum tumor necrosis factor-α (TNF-α) were assessed by ELISA. The expression of TNF-α and the cholinergic receptor, α7nAChR, in the synovial tissue was measured by RT-PCR. Nicotine treatment ameliorated cartilage destruction, promoted matrix production, reduced the serum level of TNF-α and the expression of TNF-α in the synovial tissue, and increased the expression of α7nAChR in the synovial tissue in the rat model of early stage OA. In conclusion, nicotine prevented cartilage damage and had an anti-inflammatory effect in a rat model of early stage OA. Thus nicotine may have potential as a therapeutic strategy for early stage OA.

Associations between the properties of the cartilage matrix and findings from quantitative MRI in human osteoarthritic cartilage of the knee.

The aim of this study was to investigate the associations between the properties of the cartilage matrix and the results of T2 mapping and delayed gadolinium-enhanced magnetic resonance imaging (dGEMRIC) in human knee osteoarthritic cartilage. Osteochondral samples were harvested from the middle part of the femoral condyle and tibial plateaus of 20 patients with knee osteoarthritis (OA) during total knee arthroplasty. Sagittal T2 mapping, T1pre, and T1Gd were performed using 7.0T magnetic resonance imaging (MRI). Glycosaminoglycan (GAG) distribution was evaluated by OARSI, collagen anisotropy was assessed by polarized light microscopy (PLM), and biochemical analyses measured water, GAG, and collagen content. Associations between properties of the cartilage matrix and T2 and ΔR1 (1/T1Gd-1/T1pre) values were explored using correlation analysis. T2 and ΔR1 values were significantly correlated with the degree of cartilage degeneration (OARSI grade; Ρ = 0.53 and 0.77). T2 values were significantly correlated with water content (r = 0.69; P < 0.001), GAG content (r = -0.43; P < 0.001), and PLM grade (r = 0.47; P < 0.001), but not with collagen content (r = -0.02; P = 0.110). ΔR1 values were significantly correlated with GAG content (r = -0.84; P < 0.001) and PLM grade (r = 0.41; P < 0.001). Taken together, T2 mapping and dGEMRIC results were correlated with the properties of the cartilage matrix in human knee osteoarthritic cartilage. Combination T2 mapping and dGEMRIC represents a potential non-invasive monitoring technique to detect the progress of knee OA.

Uncultured bone marrow mononuclear cells delay the dedifferentiation of unexpanded chondrocytes in pellet culture.

Uncultured bone marrow mononuclear cells (BMMC) were recently used to successfully repair damaged cartilage. However, the effect of BMMCs on the proliferation and differentiation of chondrocytes that are critical to cartilage repair is unclear. Here, we investigate the influence of BMMCs on chondrocyte dedifferentiation in pellet culture. We isolated and mixed BMMCs and chondrocytes in a 1:1 (BMMC/C) ratio and cultured in pellets (1.6 × 10(6) cells per pellet) for 2, 4, or 8 weeks. Chondrocyte differentiation was evaluated using macrography, histological examination, immunohistochemistry and gene expression analysis. While a transparent and smooth surface was observed in both BMMC/C and chondrocyte cultures over time, the former was smaller in size after 2 and 4 weeks of culture. Interestingly, after 8 weeks, BMMC/C cultures became significantly larger than chondrocyte cultures (P = 0.003). The distribution of a cartilage-specific extracellular matrix (ECM), that includes components like glycosaminoglycan (GAG) and type II collagen, was gradually reduced in chondrocyte cultures. On the other hand, while we found no obvious differences in the ECM in BMMC/C cultures between 2 and 4 weeks in vitro, after 8 weeks the concentration of ECM components decreased significantly. Further, we detected an upregulation of cartilage-specific genes in BMMC/C cultures, when compared with chondrocytes. Altogether, we demonstrate that co-culture with BMMCs delays the dedifferentiation of chondrocytes in pellet cultures in vitro. This suggests that uncultured BMMC, which can be quickly and safely obtained, could serve as a potential alternative cell source for engineering of cartilage tissue.

Three-dimensional polycaprolactone-hydroxyapatite scaffolds combined with bone marrow cells for cartilage tissue engineering.

The goal of this study was to investigate the chondrogenic potential of three-dimensional polycaprolactone-hydroxyapatite (PCL-HA) scaffolds loaded with bone marrow cells in vitro and the effect of PCL-HA scaffolds on osteochondral repair in vivo. Here, bone marrow was added to the prepared PCL-HA scaffolds and cultured in chondrogenic medium for 10 weeks. Osteochondral defects were created in the trochlear groove of 29 knees in 17 New Zealand white rabbits, which were then divided into four groups that underwent: implantation of PCL-HA scaffolds (left knee, n = 17; Group 1), microfracture (right knee, n = 6; Group 2), autologous osteochondral transplantation (right knee, n = 6; Group 3), and no treatment (right knee, n = 5; Control). Extracellular matrix produced by bone marrow cells covered the surface and filled the pores of PCL-HA scaffolds after 10 weeks in culture. Moreover, many cell-laden cartilage lacunae were observed, and cartilage matrix was concentrated in the PCL-HA scaffolds. After a 12-week repair period, Group 1 showed excellent vertical and lateral integration with host bone, but incomplete cartilage regeneration and matrix accumulation. An uneven surface of regenerated cartilage and reduced distribution of cartilage matrix were observed in Group 2. In addition, abnormal bone growth and unstable integration between repaired and host tissues were detected. For Group 3, the integration between transplanted and host cartilage was interrupted. Our findings indicate that the PCL-HA scaffolds loaded with bone marrow cells improved chondrogenesis in vitro and implantation of PCL-HA scaffolds for osteochondral repairenhanced integration with host bone. However, cartilage regeneration remained unsatisfactory. The addition of trophic factors or the use of precultured cell-PCL-HA constructs for accelerated osteochondral repair requires further investigation.

Use of quantitative MRI for the detection of progressive cartilage degeneration in a mini-pig model of osteoarthritis caused by anterior cruciate ligament transection.

BACKGROUND: To investigate the progression of cartilage degeneration using delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) and T2 mapping in a mini-pig model of osteoarthritis (OA) caused by anterior cruciate ligament transection (ACLT). METHODS: Twelve mini-pigs underwent ACLT in the left knee and were monitored by dGEMRIC (T1, gadolinium [Gd]) and T2 mapping after 2, 4, or 6 weeks (n = 4 each). No ACLT surgery was performed in four healthy control mini-pigs, which were also monitored by dGEMRIC and T2 mapping. Cartilage samples from the weight-bearing regions of the left medial femoral condyles were collected for macroscopic, histological, immunohistochemical, and biochemical analysis. Correlations between biochemical contents and T1,Gd and T2 values were evaluated using Pearson correlation analysis. RESULTS: T1,Gd values were gradually reduced and T2 values increased over time. Cartilage surfaces showed roughness at 4 weeks and additional defects at 6 weeks. Glycosaminoglycan (GAG) distribution and content gradually reduced over time (P < 0.05), and collagen distribution and anisotropy were obviously changed at 6 weeks. However, collagen content did not differ significantly among postoperative timepoints. GAG content was positively related to T1,Gd values (r = 0.888; P < 0.001) and negatively related to T2 values (r = -0.865; P < 0.001). Collagen content had no discernible correlation with T1,Gd or T2 values. CONCLUSION: dGEMRIC and T2 mapping can monitor the progression of cartilage degeneration in a mini-pig model of ACLT-induced OA, permitting early detection of OA.

Composite scaffolds composed of bone marrow mesenchymal stem cell-derived extracellular matrix and marrow clots promote marrow cell retention and proliferation.

Various biomaterials have been investigated in attempts to improve the mechanical stability of marrow clots derived from microfracture to obtain repaired tissue closely resembling hyaline cartilage. The goal of this study was to investigate the retention, adhesion, proliferation, and cartilage extracellular matrix (ECM) production of marrow clot-derived cells within a bone marrow mesenchymal stem cell-derived (BMSC-d) ECM/marrow clot composite scaffold. We fabricated BMSC-dECM/marrow clot composite scaffolds and kept them in chondrogenic medium in vitro for 1, 3, or 6 weeks. Unmodified marrow clots were used as a control. The BMSC-dECM/marrow clot composite scaffold exhibited a porous structure suitable for cell attachment and growth and further maintained cell viability. The DNA content measurements revealed that more cells proliferated in the BMSC-dECM/marrow clot composite scaffolds over time than in the marrow clots. Furthermore, the histologic, immunohistochemical, and western blot results demonstrated that the BMSC-dECM/marrow clot composite scaffold produced more hyaline-like cartilage and less fibrocartilage than the marrow clot in culture. Taken together, these findings indicate that the porous BMSC-dECM/marrow clot composite scaffold promotes the retention, attachment, and proliferation of cells from the marrow clot, and thus can stabilize the marrow clot to support chondrogenesis.

Mild degenerative changes of hip cartilage in elderly patients: an available sample representative of early osteoarthritis.

This study investigated the cellular and molecular changes which occur in cartilage from adults with femoral neck fracture (FNF) and osteoarthritis (OA), and explored the similarities in hip cartilage obtained from elderly patients and patients with early OA. Femoral heads were retrieved from 23 female patients undergoing total hip arthroplasty (THA). This group included 7 healthy patients with FNF (hFNF), 8 elderly adults with FNF (eFNF), and 8 elderly patients with hip OA (OA). After high-field MRI T2 mapping, osteochondral plugs were harvested from the weight-bearing area of femoral heads for subsequent macroscopic, histologic, and immunochemical evaluation. Additionally, the contents of cartilage matrix were analyzed, and gene expression was detected. The surface of cartilage from hFNF and eFNF patients appeared smooth, regular, and elastic, whereas it showed irregularities, thinning, and defects in OA patients. Elevated T2 values and decreased accumulation of glycosaminoglycans (GAGs) were detected in cartilage from eFNF patients. Furthermore, type I collagen accumulation was slightly increased and type X collagen concentration was obviously elevated in eFNF patients; however, type II collagen distribution and the contents and anisotropy of collagen fibrils in eFNF patients showed no significant changes. Consistent with histology and immunohistochemical results, aggrecan was downregulated and type X collagen was upregulated, while collagens types I and II showed no significant changes in eFNF patients. The cellular and molecular characteristics of hip cartilage in eFNF patients who showed no symptoms of OA were similar to those in patients with mild OA. Thus, eFNF cartilage can serve as a comparative specimen for use in studies investigating early OA.