Kristina Arvidson

A retrospective study of 236 patients with teeth restored by carbon fiber-reinforced epoxy resin posts

STATEMENT OF PROBLEM The Composipost dowel is made of stretched, aligned carbon fibres embedded in an epoxy-resin matrix. It is widely used in Europe and Canada for the restoration of endodontically treated teeth and was introduced in the United States 2 years ago as the C-Post dowel. PURPOSE This retrospective study evaluated treatment outcome of the Composipost system after 2 to 3 years. MATERIAL AND METHODS A total of 236 patients treated during a 1-year period by seven Swedish dental practitioners were included. Of those, 146 patients consented and data were collected from the dental records of the remaining patients. Thus, the material comprised 236 teeth restored with carbon fiber-reinforced epoxy resin post, 130 maxillary and 106 mandibular teeth, with a mean restoration time of 32 months (range 27 to 41). Periodontal conditions, radiographic signs, and prosthodontic results were recorded. RESULTS Five teeth (2%) had been extracted for reasons unrelated to the Composipost system. Periodontal conditions such as plaque accumulation, gingival health, bleeding on probing, and pocket depth around the teeth with Composipost dowels were similar to the control teeth. No dislodgment or root or post fractures were observed clinically or on radiographs. Radiographic examination of bone height measured from the apex to the bone margin mesially and distally showed differences on the mesial side but not on the distal surface (p < 0.05) between the Composipost-treated teeth and the controls. CONCLUSIONS Promising results after 2 to 3 years of clinical service indicate that this system can be a viable alternative to conventional post-and-core systems.

Bone regeneration and stem cells

•  Introduction •  Bone fracture healing and healing problems •  Biomaterial scaffolds and tissue engineering in bone formation ‐  Bone tissue engineering ‐  Biomaterial scaffolds ‐  Synthetic scaffolds ‐  Micro‐ and nanostructural properties of scaffolds ‐  Conclusion •  Mesenchymal stem cells and osteogenesis ‐  Bone tissue ‐  Origin of osteoblasts ‐  Isolation and characterization of bone marrow derived MSC ‐  In vitro differentiation of MSC into osteoblast lineage cells ‐  In vivo differentiation of MSC into bone ‐  Factors and pathways controlling osteoblast differentiation of hMSC ‐  Defining the relationship between osteoblast and adipocyte differentiation from MSC ‐  MSC and sex hormones ‐  Effect of aging on osteoblastogenesis ‐  Conclusion •  Embryonic, foetal and adult stem cells in osteogenesis ‐  Cell‐based therapies for bone ‐  Specific features of bone cells needed to be advantageous for clinical use ‐  Development of therapeutic biological agents ‐  Clinical application concerns ‐  Conclusion •  Platelet‐rich plasma (PRP), growth factors and osteogenesis ‐  PRP effects in vitro on the cells involved in bone repair ‐  PRP effects on osteoblasts ‐  PRP effects on osteoclasts ‐  PRP effects on endothelial cells ‐  PRP effects in vivo on experimental animals ‐  The clinical use of PRP for bone repair ‐  Non‐union ‐  Distraction osteogenesis ‐  Spinal fusion ‐  Foot and ankle surgery ‐  Total knee arthroplasty ‐  Odontostomatology and maxillofacial surgery ‐  Conclusion •  Molecular control of osteogenesis ‐  TGF‐β signalling ‐  FGF signalling ‐  IGF signalling ‐  PDGF signalling ‐  MAPK signalling pathway ‐  Wnt signalling pathway ‐  Hedgehog signalling ‐  Notch signalling ‐  Ephrin signalling ‐  Transcription factors regulating osteoblast differentiation ‐  Conclusion •  Summary

Endothelial cells influence the osteogenic potential of bone marrow stromal cells

BackgroundImproved understanding of the interactions between bone cells and endothelial cells involved in osteogenesis should aid the development of new strategies for bone tissue engineering. The aim of the present study was to determine whether direct communication between bone marrow stromal cells (MSC) and human umbilical vein endothelial cells (EC) could influence the osteogenic potential of MSC in osteogenic factor-free medium.MethodsAfter adding EC to MSC in a direct-contact system, cell viability and morphology were investigated with the WST assay and immnostaining. The effects on osteogenic differentiation of adding EC to MSC was systematically tested by the using Superarray assay and results were confirmed with real-time PCR.ResultsFive days after the addition of EC to MSC in a ratio of 1:5 (EC/MSC) significant increases in cell proliferation and cellular bridges between the two cell types were detected, as well as increased mRNA expression of alkaline phosphatase (ALP). This effect was greater than that seen with addition of osteogenic factors such as dexamethasone, ascorbic acid and β-glycerophosphate to the culture medium. The expression of transcription factor Runx2 was enhanced in MSC incubated with osteogenic stimulatory medium, but was not influenced by induction with EC. The expression of Collagen type I was not influenced by EC but the cells grown in the osteogenic factor-free medium exhibited higher expression than those cultured with osteogenic stimulatory medium.ConclusionThese results show that co-culturing of EC and MSC for 5 days influences osteogenic differentiation of MSC, an effect that might be independent of Runx2, and enhances the production of ALP by MSC.

Osteogenic Differentiation by Rat Bone Marrow Stromal Cells on Customized Biodegradable Polymer Scaffolds

In this report, poly(L-lactide-co-ε-caprolactone), poly(LLA-co-CL) and poly(L-lactide-co-1,5-dioxepan-2-one), poly(LLA-co-DXO) were evaluated and compared for potential use in bone tissue engineering constructs together with bone marrow stromal cells (BMSC). The copolymers were tailored to reduce the level of harmful tin residuals in the scaffolding. BMSC isolated from Sprague—Dawley rats were seeded onto the scaffolds and cultured in vitro for up to 21 days. Cell spreading and proliferation was analyzed after 72 h by scanning electron microscopy and thiazolyl blue tetrazolium bromide (MTT) conversion assay. Osteogenic differentiation of BMSC was evaluated by real-time PCR after 14 and 21 days of culture. Hydrophilicity was significantly different between poly(LLA-co-CL) and poly(LLA-co-DXO) with the latter being more hydrophilic. After 72 h, both scaffolds supported increased cell proliferation and the mRNA expression of osteocalcin and osteopontin was significantly increased after 21 days. Further investigation of these constructs, with lower levels of tin residuals, are being pursued.

Protein gene product 9.5-immunoreactive nerves and cells in human oral mucosa

Current conflicting information on the innervation of the human oral cavity indicates technical problems such as different detectability of the neural structures according to the various staining methods used and difficulties in reproducibility. The possibility of intraoral regional differences has not been properly considered.

In vitro and in vivo degradation profile of aliphatic polyesters subjected to electron beam sterilization

Degradation characteristics in response to electron beam sterilization of designed and biodegradable aliphatic polyester scaffolds are relevant for clinically successful synthetic graft tissue regeneration. Scaffold degradation in vitro and in vivo were documented and correlated to the macroscopic structure and chemical design of the original polymer. The materials tested were of inherently diverse hydrophobicity and crystallinity: poly(L-lactide) (poly(LLA)) and random copolymers from L-lactide and ε-caprolactone or 1,5-dioxepan-2-one, fabricated into porous and non-porous scaffolds. After sterilization, the samples underwent hydrolysis in vitro for up to a year. In vivo, scaffolds were surgically implanted into rat calvarial defects and retrieved for analysis after 28 and 91days. In vitro, poly(L-lactide-co-1,5-dioxepan-2-one) (poly(LLA-co-DXO)) samples degraded most rapidly during hydrolysis, due to the pronounced chain-shortening reaction caused by the sterilization. This was indicated by the rapid decrease in both mass and molecular weight of poly(LLA-co-DXO). Poly(L-lactide-co-ε-caprolactone) (poly(LLA-co-CL)) samples were also strongly affected by sterilization, but mass loss was more gradual; molecular weight decreased rapidly during hydrolysis. Least affected by sterilization were the poly(LLA) samples, which subsequently showed low mass loss rate and molecular weight decrease during hydrolysis. Mechanical stability varied greatly: poly(LLA-co-CL) withstood mechanical testing for up to 182 days, while poly(LLA) and poly(LLA-co-DXO) samples quickly became too brittle. Poly(LLA-co-DXO) samples unexpectedly degraded more rapidly in vitro than in vivo. After sterilization by electron beam irradiation, the three biodegradable polymers present widely diverse degradation profiles, both in vitro and in vivo. Each exhibits the potential to be tailored to meet diverse clinical tissue engineering requirements.

Toxicity of Formaldehyde to Human Oral Fibroblasts and Epithelial Cells: Influences of Culture Conditions and Role of Thiol Status

The toxicity of formaldehyde, a monomer released from certain polymeric dental materials, was studied in cultured human oral fibroblasts and epithelial cells. The influences of growth conditions were evaluated for both cell types, as well as the role of the internal and external thiol states. A one-hour exposure to formaldehyde decreased the colony-forming efficiency (CFE) of both cell types in a concentration-dependent manner, although the toxicity varied up to 100-fold with the conditions. Clearly, the presence of serum and the thiol cysteine counteracted the toxicity in fibroblasts. Similarly, pituitary extract and cysteine, or a mixture of amino acids and ethanolamines, counteracted the formaldehyde toxicity in serum-free cultures of epithelial cells. In contrast, a growth-promoting surface matrix of fibronectin and collagen did not influence the formaldehyde toxicity, as shown by both the CFE assay and a dye reduction assay. Further, a short-term change to the various growth media per se with or without the supplements serum or cysteine did not significantly alter the CFE. Analysis of the thiol state demonstrated significant differences between epithelial cells and fibroblasts, i.e., comparatively lower cellular levels of the free low-molecular-weight thiols glutathione and cysteine in fibroblasts. This result correlated to significantly higher formaldehyde toxicity in the fibroblasts than in the epithelial cells. Taken together, the results indicated the cytoprotective function of both intracellular and extracellular thiols toward formaldehyde, as well as the usefulness of thiol-free and chemically defined conditions for toxicity assessments in oral epithelial cells and fibroblasts. We conclude that the combined use of a controlled external milieu and the presumed target cell type may be advantageous in evaluations of oral toxicity mechanisms or the toxic potency of dental materials, particularly those which, like formaldehyde, may react with thiols or amines.

Growth regulation of serum-free cultures of epithelial cells from normal human buccal mucosa

SummaryHuman buccal epithelial cells have been reared from explants maintained in supplemented MCDB 153 medium. Primary epithelial outgrowths show typical structural features and uniformly express keratins; subunit analyses demonstrate expression of keratins 5, 6, 14, 16/17, and 19. The cells exhibit up to 6% colony forming efficiency and divide at about 0.8 population doublings per day on fibronectin/collagen-coated dishes at clonal density. Studies of markers of proliferation and differentiation in buccal epithelial cells indicate that epidermal growth factor, cholera toxin, retinoic acid, and pituitary extract each exhibit a distinctive ability to enhance growth and variably affect cell migration and cell surface area. Transforming growth factorβ-1 inhibits growth and increases surface area without affecting migration, involucrin expression, and cross-linked envelope formation. Moreover, exposure of cells to fetal bovine serum, the tumor promoting agent 12-O-tetradecanoylphorbol-13-acetate or an elevated Ca2+ concentration (from 0.1 to 1 mM) inhibits growth and induces squamous differentiation as indicated by inhibition of migration, increases in surface area, involucrin expression, or formation of cross-linked envelopes. The results show that epithelial cells can be reproducibly derived from explant cultures of human buccal mucosa specimens and the cells transferred under serum-free conditions. Buccal epithelial cells in culture undergo a pattern of growth and differentiation that mimics parakeratinization in vivo and variably respond to several agents shown to modulate growth of cells that originate from other types of epithelia.

Biocompatibility of Polyester Scaffolds with Fibroblasts and Osteoblast-like Cells for Bone Tissue Engineering

The aim of this study was to evaluate the in vitro cytotoxicity and cytocompatibility of the developed aliphatic polyester co-polymer scaffolds: poly(L-lactide-co-ε-caprolactone) and poly(L-lactide-co-1,5-dioxepan-2-one). The scaffolds were produced by solvent casting and particulate leaching, and tested by direct and indirect contact cytotoxicity assays on human osteoblast-like cells and mouse fibroblasts. Cell morphology was documented by light and scanning electron microscopy. Viability was assessed by the MTT, neutral red uptake, lactic dehydrogenase and apoptosis assays. Extraction tests confirmed that the scaffolds did not have a cytotoxic effect on the cells. The cells grew and spread well on the test scaffolds with good cellular attachment and viability. The scaffolds are noncytotoxic and biocompatible with the two cell types and warrant continued investigation as potential constructs for bone tissue engineering.

Effect of endothelial cells on bone regeneration using poly(L-lactide-co-1,5-dioxepan-2-one) scaffolds

Our recent in vitro study demonstrated that endothelial cells (ECs) might influence the differentiation of bone marrow stromal cells (BMSCs). Therefore, the aim of this study was to describe this effect in vivo, using a rat calvarial bone defect model. BMSCs were isolated from femurs of two-donor Lewis rats and expanded in α-minimum essential medium containing 10% fetal bovine serum. One fifth of BMSCs were induced and differentiated into ECs in an Endothelial Cell Growth Medium-2 and then characterized by a flow cytometry. The remaining BMSCs were cultured in freshly prepared osteogenic stimulatory medium, containing dexamethasone, ascorbic acid and β-glycerophosphate. Either BMSCs alone (BMSC-group) or co-cultured ECs/BMSCs (CO-group) were seeded into poly(L-lactide-co-1,5-dioxepan-2-one) [poly(LLA-co-DXO)] scaffolds, cultured in spinner flasks, and then implanted into symmetrical calvarial defects prepared in recipient rats. The animals were sacrificed after 2 months. The formation of new bone was evaluated by radiography and histology and by the expression of osteogenic markers using reverse transcriptase-polymerized chain reaction (RT-PCR). To investigate vessel formation, histological staining was performed with ECs markers. The radiographical and histological results showed more rapid bone formation in the CO- than in the BMSC-group. However, the expression of ECs marker was similar on both groups by histological analysis after 2 months postoperatively. Furthermore, the CO-group exhibited greater expression of osteogenic markers as demonstrated by RT-PCR. The results are consistent with the previous in vitro findings that poly(LLA-co-DXO) scaffold might be suitable candidate for bone tissue engineering. In vivo, bone regeneration was enhanced by a construct of the polymer scaffold loaded with co-cultured cells.