This study investigates the impact of hydroxyapatite (HA) nanoparticles (NPs) on the cellular responses of poly(L-lactide-co-ε-caprolactone) (PLCL) scaffolds in bone tissue engineering applications. Three types of PLCL scaffolds were fabricated, varying in HANPs content. Saos-2 osteoblast-like cells (OBs) and THP-1-derived osteoclast-like cells (OCs) were co-cultured on the scaffolds, and cell proliferation was assessed using the MTS assay. The amount of double-stranded DNA (dsDNA) was quantified to evaluate cell proliferation. Expression levels of OBs and OCs markers were analyzed via quantitative polymerase chain reaction (qPCR) and the production of Collagen type I was visualized using confocal microscopy. Additionally, enzymatic activity of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP or ACP5) was measured to assess OB and OC function, respectively. Interestingly, despite the scaffold's structured character supporting the growth of the Saos-2 OBs and THP-1-derived OCs coculture, the incorporation of HANPs did not significantly enhance cellular responses compared to scaffolds without HANPs, except for collagen type I production. These findings suggest the need for further investigation into the potential benefits of HANPs in bone tissue engineering applications. Nevertheless, our study contributes valuable insights into optimizing biomaterial design for bone tissue regeneration, with implications for drug screening and material testing protocols.

• Klíčová slova
• PLCL, bone regeneration, hydroxyapatite, osteoblasts, osteoclasts, scaffold, tissue engineering,
• MeSH
• buněčné linie MeSH
• hydroxyapatit * chemie   MeSH
• kokultivační techniky MeSH
• lidé MeSH
• nanočástice * chemie   MeSH
• nanovlákna * chemie   MeSH
• osteoblasty cytologie   metabolismus   účinky léků   MeSH
• osteogeneze MeSH
• osteoklasty cytologie   metabolismus   účinky léků   MeSH
• polyestery * chemie   MeSH
• proliferace buněk účinky léků   MeSH
• regenerace kostí * účinky léků   MeSH
• THP-1 buňky MeSH
• tkáňové inženýrství metody   MeSH
• tkáňové podpůrné struktury chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hydroxyapatit * MeSH
• poly(lactic acid-co-epsilon-caprolactone) MeSH Prohlížeč
• polyestery * MeSH

Extracellular vesicles can play an important role in the processes occurring after stem cell transplantation, preventing cell apoptosis, stimulating immunological processes, and promoting the synthesis of extracellular matrix. Human follicular fluid (FF) can be a source of a subpopulation of cells with mesenchymal stem cells (MSCs) properties. Moreover these subpopulations of FF cells can differentiate into osteoblasts. In presented studies flow cytometry of ovarian FF cells confirmed positive expression of MSCs markers such as: CD44, CD90, CD105, CD73 and negative expression of a hematopoietic marker: CD45. The CD90+, CD105+, CD45- cell subpopulation has been obtained during magnetic separation using appropriate antibodies conjugated with microbeads. The extracellular vesicles (EVs) secreted by the cells during osteodifferentiation process differed from those secreted by cells culture in the basal medium. Based on the previous and current electron microscopy research, changes in size, number, and shape would support the notion that released EVs could be crucial to the ovarian FF cell subpopulation differentiation process. Osteogenic differentiation has been confirmed via Alizarin red staining. Therefore, follicular fluid (FF) can be a new source of a cell subpopulation with MSC properties, with the cells capable of differentiating into the osteogenic lineage. EVs could play a key role as mediators in tissue regeneration, especially bone tissue regeneration.

• Klíčová slova
• Extracellular vesicles, Human granulosa cells, Osteodifferentiation,
• MeSH
• buněčná diferenciace * MeSH
• extracelulární vezikuly * ultrastruktura   metabolismus   MeSH
• folikulární tekutina * cytologie   metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky * cytologie   metabolismus   MeSH
• osteoblasty cytologie   metabolismus   MeSH
• osteogeneze * MeSH
• průtoková cytometrie MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The etiology of bone loss in celiac disease (CeD) remains a clinical challenge, with uncertainties present such as the extent of involvement of malabsorption and inflammation-induced osteoresorption processes in development of osteopenia/osteoporosis (OPN/OP), or reasons for failure to achieve healthy bone mass (BMD) even after long-term gluten-free diet (GFD) treatment. This observational prospective study explores the in vitro osteoclastogenic potential of peripheral blood precursors originating from adult active (newly diagnosed and untreated) celiac disease patients (aCeD) and describes the longitudinal changes in osteoclastogenesis after long-term adherence to GFD. To find connections between in vitro observations and in vivo bone metabolism changes, serum levels of 25(OH)D3, PTH, bCTX, PINP, CRP, IL-6, RANKL and OPG were measured before and after GFD and levels of these markers were correlated with the rate of osteoclastogenesis in vitro. OPG and IL-6 showed associations with BMD and/or presence of OPN/OP. Patients after GFD (CeD-GFD) exhibited improved BMD and increased serum 25(OH)D3 levels, alongside reduced bCTX and PINP levels. Compared to healthy donors, aCeD osteoclast genesis in vitro was higher and, surprisingly, remained elevated even in CeD-GFD patients. Negative correlation was found between osteoclastogenesis rate and serum OPG in aCeD, while osteoclastogenesis rate positively correlated with PTH in CeD-GFD. These results highlight OPG as marker for risk of OPN/OP in CeD and suggest that improvement of BMD after GFD is a result of uncoupling between bone metabolism and osteoresorptive action of osteoclasts after GFD.

• Klíčová slova
• Bone metabolism, Celiac disease, Gluten-free diet, Osteoclastogenesis, Osteoporosis,
• MeSH
• bezlepková dieta * MeSH
• celiakie * dietoterapie   komplikace   krev   patofyziologie   MeSH
• dospělí MeSH
• interleukin-6 * krev   MeSH
• kosti a kostní tkáň * metabolismus   MeSH
• kostní denzita MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• osteogeneze * MeSH
• osteoklasty metabolismus   MeSH
• osteoprotegerin * krev   MeSH
• parathormon krev   MeSH
• prospektivní studie MeSH
• vitamin D * krev   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• Názvy látek
• IL6 protein, human MeSH Prohlížeč
• interleukin-6 * MeSH
• osteoprotegerin * MeSH
• parathormon MeSH
• TNFRSF11B protein, human MeSH Prohlížeč
• vitamin D * MeSH

OBJECTIVE: Insulin-sensitizing drugs, despite their broad use against type 2 diabetes, can adversely affect bone health, and the mechanisms underlying these side effects remain largely unclear. Here, we investigated the different metabolic effects of a series of thiazolidinediones, including rosiglitazone, pioglitazone, and the second-generation compound MSDC-0602K, on human mesenchymal stem cells (MSCs). METHODS: We developed 13C subcellular metabolomic tracer analysis measuring separate mitochondrial and cytosolic metabolite pools, lipidomic network-based isotopologue models, and bioorthogonal click chemistry, to demonstrate that MSDC-0602K differentially affected bone marrow-derived MSCs (BM-MSCs) and adipose tissue-derived MSCs (AT-MSCs). In BM-MSCs, MSDC-0602K promoted osteoblastic differentiation and suppressed adipogenesis. This effect was clearly distinct from that of the earlier drugs and that on AT-MSCs. RESULTS: Fluxomic data reveal unexpected differences between this drug's effect on MSCs and provide mechanistic insight into the pharmacologic inhibition of mitochondrial pyruvate carrier 1 (MPC). Our study demonstrates that MSDC-0602K retains the capacity to inhibit MPC, akin to rosiglitazone but unlike pioglitazone, enabling the utilization of alternative metabolic pathways. Notably, MSDC-0602K exhibits a limited lipogenic potential compared to both rosiglitazone and pioglitazone, each of which employs a distinct lipogenic strategy. CONCLUSIONS: These findings indicate that the new-generation drugs do not compromise bone structure, offering a safer alternative for treating insulin resistance. Moreover, these results highlight the ability of cell compartment-specific metabolite labeling by click reactions and tracer metabolomics analysis of complex lipids to discover molecular mechanisms within the intersection of carbohydrate and lipid metabolism.

• Klíčová slova
• Adipocyte, Bone marrow, Lipid flux analysis, Mitochondrial pyruvate carrier, Tracer metabolomics,
• MeSH
• adipogeneze * účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• hypoglykemika * farmakologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• metabolomika metody   MeSH
• mezenchymální kmenové buňky účinky léků   metabolismus   MeSH
• osteogeneze * účinky léků   MeSH
• pioglitazon MeSH
• rosiglitazon farmakologie   MeSH
• thiazolidindiony * farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hypoglykemika * MeSH
• pioglitazon MeSH
• rosiglitazon MeSH
• thiazolidindiony * MeSH

INTRODUCTION: The primary aim of this study was to assess the amount and long-term stability of orthodontically created bone in patients with agenesis of maxillary lateral incisors after canine distalization. The secondary aim was to explore the impact of patient age on the process of alveolar bone resorption. METHODS: A group of patients with agenesis of the maxillary permanent lateral incisor was examined at 4 time points: the beginning of orthodontic treatment (T1, n = 80), the end of treatment (T2, n = 80), 2-5 years after treatment (T3, n = 79), and 12-15 years after treatment (T4, n = 32). The width of the edentulous alveolar bone was measured from study casts at the level of the bone ridge (point A) and 5 mm apically from the alveolar ridge (point B). Alveolar ridge height was also recorded using panoramic radiographs at all time points. Paired t tests, 2-sample t tests, Friedman test with Bonferroni correction, Spearman`s correlation, and linear regression tests were used to analyze the data. RESULTS: The alveolar ridge width was reduced by an average of 0.44 mm at point A and 0.47 mm at point B during the 12-15 years after treatment (T2-T4) and by 0.21 mm and 0.19 mm during the last 10 years (T3-T4). The alveolar ridge height was reduced by 0.59 mm between T2 and T4 and by 0.05 mm between T3 and T4. All reductions in ridge width and height were statistically significant (P <0.05). However, no significant correlation was observed between patient age and changes in alveolar bone parameters (P >0.05). CONCLUSIONS: Although the reductions in alveolar ridge dimensions were statistically significant, the orthodontically created bone after canine distalization remained stable 12-15 years after treatment in both the horizontal and vertical dimensions. Patient age did not significantly influence alveolar bone changes.

• MeSH
• anodoncie * terapie   MeSH
• čelist bezzubá * diagnostické zobrazování   MeSH
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• maxila * patologie   diagnostické zobrazování   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• osteogeneze * fyziologie   MeSH
• posun zubů * metody   MeSH
• processus alveolaris * patologie   diagnostické zobrazování   MeSH
• rentgendiagnostika panoramatická MeSH
• resorpce alveolární kosti etiologie   diagnostické zobrazování   MeSH
• řezáky * abnormality   MeSH
• věkové faktory MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Treatment of gingival fibroblasts with PDL extracellular vesicles results in promotion of Wnt signalling pathway and osteogenic differentiation. PDL secretome shows selective wound healing and matrix remodelling which can have implications for future periodontal regenerative strategies.

• MeSH
• buněčná diferenciace MeSH
• extracelulární vezikuly * fyziologie   MeSH
• fibroblasty fyziologie   MeSH
• gingiva cytologie   MeSH
• hojení ran fyziologie   MeSH
• lidé MeSH
• osteogeneze fyziologie   MeSH
• periodontální vaz * cytologie   fyziologie   MeSH
• regenerace * fyziologie   MeSH
• signální dráha Wnt fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

This study introduces the first fabrication of porous titanium/hydroxyapatite interpenetrating phase composites through an innovative processing method. The approach combines additive manufacturing of a customized titanium skeleton with the infiltration of an injectable hydroxyapatite foam, followed by in situ foam hardening at physiological temperature. This biomimetic process circumvents ceramic sintering and metal casting, effectively avoiding the formation of secondary phases that can impair mechanical performance. Hydroxyapatite foams, prepared using two foaming agents (polysorbate 80 and gelatine), significantly reinforce the titanium skeleton while preserving the microstructural characteristics essential for osteoinductive properties. The strengthening mechanisms rely on the conformation of the foams to the titanium surface, thereby enabling stable mechanical interlocking and effective interfacial stress transfer. This, combined with the mechanical constriction of phases, enhances damage tolerance and mechanical reliability of the interpenetrating phase composites. In addition, the interpenetrating phase composites feature a network of concave pores with an optimal size for bone repair, support human osteoblast proliferation, and exhibit mechanical properties compatible with bone, offering a promising solution for the efficient and personalized reconstruction of large bone defects. The results demonstrate a significant advancement in composite fabrication, integrating the benefits of additive manufacturing for bone repair with the osteogenic capacity of calcium phosphate ceramics.

• Klíčová slova
• Bone repair, Ceramic matrix composite, Hydroxyapatite, Infiltration, Self-hardening, Titanium,
• MeSH
• biokompatibilní materiály chemie   farmakologie   MeSH
• hydroxyapatit * chemie   farmakologie   MeSH
• kosti a kostní tkáň účinky léků   MeSH
• kostní náhrady chemie   farmakologie   MeSH
• lidé MeSH
• osteoblasty účinky léků   MeSH
• osteogeneze účinky léků   MeSH
• poréznost MeSH
• proliferace buněk účinky léků   MeSH
• regenerace kostí účinky léků   MeSH
• testování materiálů MeSH
• titan * chemie   farmakologie   MeSH
• tkáňové podpůrné struktury chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biokompatibilní materiály MeSH
• hydroxyapatit * MeSH
• kostní náhrady MeSH
• titan * MeSH

Craniofacial morphogenesis depends on complex cell fate decisions during the differentiation of post-migratory cranial neural crest cells. Molecular mechanisms of cell differentiation of mesenchymal cells to developing bones, cartilage, teeth, tongue, and other craniofacial tissues are still poorly understood. We performed single-cell transcriptomic analysis of craniofacial mesenchymal cells derived from cranial NCCs in mouse embryo. Using FACS sorting of Wnt1-Cre2 progeny, we carefully mapped the cell heterogeneity in the craniofacial region during the initial stages of cartilage and bone formation. Transcriptomic data and in vivo validations identified molecular determinants of major cell populations involved in the development of lower and upper jaw, teeth, tongue, dermis, or periocular mesenchyme. Single-cell transcriptomic analysis of Meis2-deficient mice revealed critical gene expression differences, including increased osteogenic and cell adhesion markers. This leads to affected mesenchymal cell differentiation and increased ossification, resulting in impaired bone, cartilage, and tongue formation.

• Klíčová slova
• Bone, Cartilage, Craniofacial development, Meis2, Neural crest,
• MeSH
• analýza genové exprese jednotlivých buněk MeSH
• chondrogeneze genetika   MeSH
• crista neuralis cytologie   embryologie   MeSH
• homeodoménové proteiny genetika   MeSH
• kraniofaciální abnormality * embryologie   genetika   MeSH
• mezenchymální kmenové buňky metabolismus   MeSH
• myši knockoutované MeSH
• myši MeSH
• osteogeneze * genetika   MeSH
• sekvenování transkriptomu MeSH
• vývojová regulace genové exprese * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• homeodoménové proteiny MeSH
• Mrg1 protein, mouse MeSH Prohlížeč

Caspase-12 is a molecule whose functions are still not well understood. Although its expression has been found in various tissues, specific roles have been described in only a few cases. These include the effect of caspase-12 on murine bone cell differentiation during craniofacial development. This work focused on the development of the limbs taking place through endochondral ossification, which precedes the formation of the cartilaginous growth plate. Caspase-12 was described here for the first time in growth plate chondrocytes during physiological development. Using pharmacological inhibition, caspase-12 was found to affect chondrogenesis. Limb-derived micromass cultures showed a significantly increased area of chondrogenic nodules after caspase-12 inhibition and there were changes in gene expression, the most significant of which was the reduction of Mmp9. These data point to potential new functions of caspase-12 in chondrogenesis.

• Klíčová slova
• Caspase-12, Chondrocyte, Chondrogenesis, Differentiation, Growth plate,
• MeSH
• buněčná diferenciace MeSH
• chondrocyty * MeSH
• chondrogeneze * fyziologie   MeSH
• inhibitory kaspas farmakologie   MeSH
• kaspasa 12 * metabolismus   genetika   MeSH
• kultivované buňky MeSH
• matrixová metaloproteinasa 9 metabolismus   genetika   MeSH
• myši MeSH
• růstová ploténka růst a vývoj   MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• inhibitory kaspas MeSH
• kaspasa 12 * MeSH
• matrixová metaloproteinasa 9 MeSH

Bone nonunion delays fracture end repair and is associated with inflammation. Although bone nonunion can be effectively repaired in clinical practice, many cases of failure. Studies have confirmed that BMP-2 and nHA/PA66 repaired bone defects successfully. There are few studies on the effects of the combined application of BMP-2 and NHA/PA66 on bone nonunion osteogenesis and inflammation. We aimed to investigate the expression level of inflammation-related genes in patients with bone nonunion and the effect of BMP-2-infected mesenchymal stem cells combined with nHA/PA66 on the level of inflammation in femur nonunion rats. We searched for a gene expression profile related to bone nonunion inflammation (GSE93138) in the GEO public database. Bone marrow mesenchymal stem cells (MSCs) of SD rats were cultured and passed through. We infected the third generation of MSCs with lentivirus carrying BMP-2 and induced the infected MSCs to bone orientation. We detected the expression level of BMP-2 by RT-PCR and the cell viability and alkaline phosphatase (ALP) activity by CCK8 and then analyzed the cell adhesion ability. Finally, the levels of related inflammatory factors, including C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and Erythrocyte Sedimentation Rate (ESR), were detected in nonunion rats. Our findings: The patients with nonunion had up-regulated expression of 26 differentially inflammatory genes. These genes are mainly enriched in innate immune response, extracellular region, calcium ion binding, Pantothenate and CoA biosynthesis pathways. The expression level of BMP-2 in the Lenti-BMP-2 group was higher (vs. empty lentivirus vector group: t=5.699; vs. uninfected group t=3.996). The cell activity of the MSCs + BMP-2 + nHA/PA66 group increased gradually. After being combined with nHA/PA66, MSCs transfected with BMP-2 spread all over the surface of nHA/PA66 and grew into the material pores. MSCs + BMP-2 + nHA/PA66 cells showed positive ALP staining, and the OD value of ALP was the highest. The levels of CRP, IL-6, TNF-alpha, and ESR in the MSCs + BMP-2 + nHA/PA66 group were lower than those in the MSCs and MSCs + nHA/PA66 group but higher than those in MSCs + BMP-2 group. The above comparisons were all P<0.05. The findings demonstrated that the expression level of inflammation-related genes increased in the patients with bone nonunion. The infection of MSCs by BMP-2 could promote the directed differentiation of MSCs into osteoblasts in the bone marrow of rats, enhance the cell adhesion ability and ALP activity, and reduce inflammation in rats with bone nonunion.

• MeSH
• dospělí MeSH
• femur metabolismus   patologie   MeSH
• fraktury femuru metabolismus   genetika   MeSH
• kostní morfogenetický protein 2 * metabolismus   genetika   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• mezenchymální kmenové buňky * metabolismus   MeSH
• nezhojené fraktury * genetika   metabolismus   MeSH
• osteogeneze MeSH
• potkani Sprague-Dawley * MeSH
• transplantace mezenchymálních kmenových buněk * MeSH
• zánět * metabolismus   genetika   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• krysa rodu Rattus MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• BMP2 protein, human MeSH Prohlížeč
• kostní morfogenetický protein 2 * MeSH