The design and application of an individualized 3D printing assisted guide plates in assisting sacroiliac screws insertion.

OBJECTIVE: Currently, the sacroiliac screws insertion still faces several challenges in the fixation of pelvic and acetabular injuries. This study was aimed to design a personalized three-dimensional (3D) printing assisted guide plates to assist sacroiliac screws insertion, so as to provide a reference for further clinical applications. METHODS: Eight pelvic specimens (5 males and 3 females) of normal adults were used to simulate actual operation. After thin-layer CT scanning, the 3D models of pelvis were established based on the images data. Furthermore, in Mimics 17.0 software, the screw entry points and screw channels of sacroiliac screws were further simulated and designed, and the appropriate range of the posterior superior iliac spine was selected to establish and print the virtual guide plates. Then, the simulated screws insertion was performed in vitro, the pelvic specimens after screws insertion were scanned again by CT, and the effect of screws insertion was further evaluated. RESULTS: A total of 16 sacroiliac screw guide plates were designed and printed, and 48 screws were inserted on both sides. Therein, 45 screws were completely located in the sacral vertebra, which was determined as grade 0, with an accuracy rate of 93.2%. The other 3 screws penetrated the anterior cortex or sacral canal of sacral vertebra, including 2 screws in Grade 1 (4.1%) and 1 screw in Grade 2 (2.1%). Compared with the simulated screw channels, the anterior and posterior offset angles of the cross section were (0.912 ± 0.625) ° and (0.802 ± 0.681) ° respectively, with no significant difference (p > 0.05). The upper and lower offset angles of coronal plane were (1.158 ± 0.823) ° and (1.034 ± 0.908) ° respectively, and there was no significant difference (p > 0.05). CONCLUSIONS: 3 D printing guide plates assisted sacroiliac screws insertion can enhance the stability of pelvic posterior ring fixation and assist surgeons to reduce the difficulty of operation.

Bone mesenchymal stem cells (BMSCs)-derived exosomal microRNA-21-5p regulates Kruppel-like factor 3 (KLF3) to promote osteoblast proliferation in vitro.

Bone mesenchymal stem cells (BMSCs)-derived exosomes (Exos) play important roles in osteoporosis, while the regulation of microRNA (miR)-21-5p remains unclear. The BMSCs-derived exosomes were isolated from femoral bone marrow of trauma patients, which were then used to stimulate human osteoblasts (hFOB1.19 cells). The miR-21-5p mimic or inhibitor was transfected into BMSCs to overexpress or knockdown miR-21-5p. The functions of miR-21-5p in osteoporosis were assessed by cell counting kit-8 (CCK-8) assay, alkaline phosphatase (ALP) staining and alizarin red staining assays. We found that BMSCs-derived exosomes could enhance proliferation, osteoblastic differentiation and ALP activity of hFOB1.19 cells. BMSCs-derived exosomes with upregulated miR-21-5p could further enhance these protective impacts compared with that in BMSCs-derived exosomes, while BMSCs-derived exosomes with downregulated miR-21-5p reduced these cell phenotypes. MiR-21-5p could directly bind to the 3'-untranslated region (UTR) of Kruppel-like factor 3 (KLF3), and knockdown of KLF3 obviously attenuated these inhibitory effects of BMSCs-derived exosomes with downregulated miR-21-5p on osteoblastic differentiation and ALP activity of hFOB1.19 cells. In summary, BMSCs-derived exosomal miR-21-5p improved osteoporosis through regulating KLF3, providing a potential therapeutic strategy for osteoporosis.

The Novel Application of Three-Dimensional Printing Assisted Patient-Specific Instrument Osteotomy Guide in the Precise Osteotomy of Adult Talipes Equinovarus.

OBJECTIVE: This current research is aimed at assessing clinical efficacy and prognosis of three-dimensional (3D) printing assisted patient-specific instrument (PSI) osteotomy guide in precise osteotomy of adult talipes equinovarus (ATE). METHODS: We included a total of 27 patients of ATE malformation (including 12 males and 15 females) from June 2014 to June 2018 in the current research. The patients were divided into the routine group (n = 12) and 3D printing group (n = 15) based on different operative methods. The parameters, including the operative time, intraoperative blood loss, complications, time to obtain bony fusion, functional outcomes based on American Orthopedic Foot and Ankle Society (AOFAS), and International Congenital Clubfoot Study group (ICFSG) scoring systems between the two groups were observed and recorded regularly. RESULTS: The 3D printing group exhibits superiorities in shorter operative time, less intraoperative blood loss, higher rate of excellent, and good outcomes presented by ICFSG score at last follow-up (P < 0.001, P < 0.001, P = 0.019) than the routine group. However, there was no significant difference exhibited in the AOFAS score at the last follow-up and total rate of complications between the two groups (P = 0.136, P = 0.291). CONCLUSION: Operation assisted by 3D printing PSI osteotomy guide for correcting the ATE malformation is novel and feasible, which might be an effective method to polish up the precise osteotomy of ATE malformation and enhance the clinical efficacy.

MicroRNA-197-3p Inhibits the Osteogenic Differentiation in Osteoporosis by Down-Regulating KLF 10.

BACKGROUND: Studies have shown that microRNA (miRNA) regulates gene expression of osteoporosis (OS). It is known that miR-197-3p is abnormally expressed in osteoporosis. This study is to investigate the mechanism of miR-197-3p in regulating osteoblast differentiation. METHODS: Rats were ovariectomized to establish an animal model of postmenopausal osteoporosis. The expression of miR-197-3p and KLF10 was detected in ovariectomized rat models. Primary osteoblasts and MC3T-E1 cells were divided into the control group, miR-197-3p inhibitor group, NC inhibitor group and miR-197-3p inhibitor + si-KLF10 group. The expression of miR-197-3p and Kruppel-like factor 10 (KLF10) was detected by qRT-PCR and Western blot. The relationship between miR-197-3p and KLF10 was analyzed by bioinformatics and luciferase reporter assay. Cell viability was evaluated by MTT assay. The ALP activity measurement and mineralization analysis were performed. RESULTS: The expression of miR-197-3p was significantly raised in ovariectomized osteoporosis rats. During the differentiation of osteoblasts, the expression of miR-197-3p was significantly decreased, while the expression of KLF10 was significantly raised in primary osteoblasts and MC3E3T1 cells. The expression of RUNX2, ALP, OCN and OSX in miR-197-3p inhibitor group and MC3T3-E1 group was significantly raised, and the cell survival rate and mineralized nodule were raised as well. KLF10 may be the downstream target gene of miR-197-3p. After co-transfection of miR-197-3p inhibitor and si-klf10, ALP, Runx2, OCN and OSX mRNA, cell survival rate and mineralized nodule were significantly decreased in primary osteoblasts and MC3T3-E1 cells. CONCLUSION: MiR-197-3p Inhibition promoted osteoblast differentiation and reduced OS by up-regulating KLF10.

Dioscin stimulates differentiation of mesenchymal stem cells towards hypertrophic chondrocytes in vitro and endochondral ossification in vivo.

Dioscin has been shown to play important roles in suppression of osteoclast maturation. It is proposed as a potential natural product for the treatment of osteoclast-related diseases. We hypothesized in this study that treatment of dioscin on bone marrow mesenchymal stem cells (BMSCs) could increase the osteo-chondrogenic differentiation of BMSCs and promote endochondral ossification of BMSCs in bone fracture environment. BMSCs were extracted from femur and tibia of male C57b mice. Stemness of BMSCs was studied by performing proliferation assay and multilineage differentiation. Glycosaminoglycans (GAG) and collagen contents were assessed to examine the chondrogenesis of BMSCs. Real time quantitative PCR was carried out to examine the expression of hypertrophic marker collagen type X. Efficacy of Dioscin was then tested in mouse bone fracture model on the distal side of femur. Results showed treatment of dioscin on BMSCs increased chondrogenic differentiation of BMSCs as well as the expression of collagen type X. Local delivery of dioscin promoted endochondral ossification at bone fractured site, as shown by histological examination. Results of immunohistochemistry showed that dioscin increased collagen type X expression in bone facture model of mice. In conclusion, our results demonstrated that treatment of dioscin promote the hypertrophic differentiation of BMSCs derived chondrocytes. Dioscin could be a useful drug to promote bone regeneration after fracture.

[A finite element analysis of pelvic ring reconstruction with fibular autograft following periacetabular tumor resection].

OBJECTIVE: To establish sophisticated three-dimensional finite element model of reconstructing the whole pelvis and defects in pelvis caused by the resection of periacetabular tumor, and to research the stress distribution regularity of the pelvis reconstructed by the fibular transplantation through three different internal fixation techniques. METHODS: The CT datasets including L3 to middle-femur, unilateral fibular and internal fixation system from 1 healthy 35-year-old male volunteer were collected to establish finite element models of reconstructing the pelvis after the resection of periacetabular tumors through 3 different internal fixation means, namely fibular with plates, pedicle-rods and sacral-iliac rods. Bilateral leg standing position was simulated, then vertical load of 500 N was imposed on the surface of L3, the stress distribution regularity of reconstructed pelvis, transplanted fibular and internal fixation system were evaluated. RESULTS: The finite element models of the pelvis reconstruction after resection of periacetabular tumors were established. The stress concentration of transplanted fibular was extremely high in the vicinity of the host junction sites. For the three internal fixation systems, the connection between steel plate and screw or between titanium bar and screw inclined to have stress concentration; and when the titanium bar was adopted to reconstruct, the transplanted fibular and the healthy side of femoral neck had less stress concentration, while sacral-iliac rods had the most obvious stress concentration. CONCLUSION: For the reconstruction pelvis, the three fibula transplantation and steel plate internal fixation are consistent with intact state of pelvis in terms of the stress distribution, which is a relatively good method for the treatment of bone defect after periacetabular tumor. The finite element model can be used as a tool for the pelvis biomechanics research.

Venous drainage in retrograde island flap: an experimental study using fluorescence tracing technique.

The pathway of venous drainage in retrograde island flaps was investigated by fluorescence tracing technique using the saphenous fasciocutaneous flap in New Zealand White rabbits. Forty animals were allocated into four groups according to the different times at 30 minutes (I), 24 hours (II), 72 hours (III), and 7 days (IV) after the operation. According to the different routes to give tracer, each group was further allocated into two subgroups of the artery injection and vein injection. For each animal, one hindlimb was assigned as the experimental side, the contralateral side as control without giving tracer. The erythrocytes were separated, labeled with fluorescein isothiocyanate (FITC), detected, and injected into the artery or vein. Subsequently, the flaps were harvested 5 seconds after injection and immediately frozen, sectioned, and observed under microscope. In group I and II, the fluorescence was observed mainly around the vessel adventitia of the vein and artery and tunica intima of the artery. In group III, there was weak fluorescence observed in the lumen of vein. In group IV, fluorescence was distributed principally in the lumen of the vein. In addition, fluorescence was not observed in the saphenous nerve in group I and there was mild fluorescence in the saphenous nerve in groups II, III, and IV. These findings suggest that the venous return is through "bypass route" in earlier period. In later period, the venous retrograde return is through "bypass route" and "incompetent valves route;" however, "incompetent valves route" becomes the main route.

[Study of venous drainage in retrograde island flaps by fluorescence tracing technique in a rabbit model].

OBJECTIVE: To investigate the venous drainage in retrograde island flaps by fluorescence tracing technique and to observe the pathway of venous drainage. METHODS: The 0.1 mL venous blood was collected from the marginal ear vein of every rabbit (n=20), respectively, and erythrocytes were separated by centrifugation and then were labeled with FITC. Positive rate and fluorescence intensity of FITC-labeled RBC were detected by flow cytometry. RBC amorphous was observed under the inverted fluorescence microscope. Saphenous retrograde island fasciocutaneous flap and antegrade island fasciocutaneous flap (4.0 cm x 3.0 cm in size with vascular pedicle length of 3.0 cm) were successfully established in hind limbs of 20 New Zealand white rabbits. One hind limb of each rabbit was randomly assigned as the experimental group and the contralateral side was assigned as the control. The same flap was established in the control group without any fluorescence tracer. According to retrograde or antegrade flaps, the experimental group was divided into 2 groups with 10 rabbits in each group. And then, according to different pathways of tracer-giving, each group was divided into 2 subgroups of artery and vein, with 5 rabbits in each subgroup. The labeled erythrocytes (5 microL) were injected into artery or vein and then flaps were cut down 5 seconds later. The flaps were immediately frozen and chipped (5-7 microm). Consecutive three frozen sections were made and two of them were stained with HE and GENMED, respectively, but the third one was squashed without staining. All frozen sections were observed under the microscope. RESULTS: Positive rate of FITC-labeled RBC was beyond 99% and fluorescence intensity was more than or equal to 10(3). FITC-labeled RBC showed steady green fluorescence under the inverted fluorescence microscope. Fluorescence appeared in all experimental groups, but none was found in the control groups. In antegrade island flap group, fluorescence appeared mainly in lumen of vein, wall of vein and inner membrane and outer membrane of artery. In retrograde island flap group, fluorescence distributed principally in inner membrane and outer membrane of artery and wall of vein. CONCLUSION: The fluorescence tracing is applicable to the research of venous drainage. Venous drainage in the antegrade island flaps is mainly through lumen of vein, wall of vein and inner membrane and outer membrane of artery. While, venous drainage in retrograde island flaps is principally through inner membrane and outer membrane of artery and wall of vein.