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Characterization of bone marrow mononuclear cells on biomaterials for bone tissue engineering in vitro.

Henrich D, Verboket R, Schaible A, Kontradowitz K, Oppermann E, Brune JC, Nau C, Meier S, Bonig H, Marzi I, Seebach C - Biomed Res Int (2015)

Bottom Line: On day 14, metabolic activity was significantly increased in BMC seeded on DBM in comparison to BMC seeded on BS.Likewise increased VEGF-synthesis was observed on day 2 in BMC seeded on DBM when compared to BMC seeded on BS.Beta-TCP and DBM were similar and both superior to BS, suggesting either as suitable materials for spatial restriction of BMC used for regenerative medicine purposes in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Trauma, Hand and Reconstructive Surgery, Hospital of the Goethe University, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany.

ABSTRACT
Bone marrow mononuclear cells (BMCs) are suitable for bone tissue engineering. Comparative data regarding the needs of BMC for the adhesion on biomaterials and biocompatibility to various biomaterials are lacking to a large extent. Therefore, we evaluated whether a surface coating would enhance BMC adhesion and analyze the biocompatibility of three different kinds of biomaterials. BMCs were purified from human bone marrow aspirate samples. Beta tricalcium phosphate (β-TCP, without coating or coated with fibronectin or human plasma), demineralized bone matrix (DBM), and bovine cancellous bone (BS) were assessed. Seeding efficacy on β-TCP was 95% regardless of the surface coating. BMC demonstrated a significantly increased initial adhesion on DBM and β-TCP compared to BS. On day 14, metabolic activity was significantly increased in BMC seeded on DBM in comparison to BMC seeded on BS. Likewise increased VEGF-synthesis was observed on day 2 in BMC seeded on DBM when compared to BMC seeded on BS. The seeding efficacy of BMC on uncoated biomaterials is generally high although there are differences between these biomaterials. Beta-TCP and DBM were similar and both superior to BS, suggesting either as suitable materials for spatial restriction of BMC used for regenerative medicine purposes in vivo.

No MeSH data available.


Related in: MedlinePlus

Seeding efficacy of BMC depending on the surface coating and medium conditions. BMCs (n = 16) were seeded on β-TCP either coated with fibronectin or human fresh frozen plasma and on uncoated β-TCP in the presence of MesenCult medium or DMEM. Seeding efficacy (%) is plotted on the ordinate. No significant differences in seeding efficacy were observed (a). Representative fluorescence microscopy images of DAPI stained BMC seeded under the different conditions are shown in (b). Similar amounts of adhering BMC were seen under each condition. No selective enrichment or depletion of specific progenitor cell subpopulation was observed on the surface of all biomaterials in various medium conditions as shown in (c). The percentages of progenitor cell types which resemble EPC (CD34+/CD45+; CD34+/CD133+/CD45+) and MSC (CD45−/CD271+) were analyzed in the fraction of nonadhering cells. The similar values indicate that no selective enrichment of progenitor cell subpopulations occurred due to the different culture and surface conditions.
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fig1: Seeding efficacy of BMC depending on the surface coating and medium conditions. BMCs (n = 16) were seeded on β-TCP either coated with fibronectin or human fresh frozen plasma and on uncoated β-TCP in the presence of MesenCult medium or DMEM. Seeding efficacy (%) is plotted on the ordinate. No significant differences in seeding efficacy were observed (a). Representative fluorescence microscopy images of DAPI stained BMC seeded under the different conditions are shown in (b). Similar amounts of adhering BMC were seen under each condition. No selective enrichment or depletion of specific progenitor cell subpopulation was observed on the surface of all biomaterials in various medium conditions as shown in (c). The percentages of progenitor cell types which resemble EPC (CD34+/CD45+; CD34+/CD133+/CD45+) and MSC (CD45−/CD271+) were analyzed in the fraction of nonadhering cells. The similar values indicate that no selective enrichment of progenitor cell subpopulations occurred due to the different culture and surface conditions.

Mentions: The seeding efficacy was generally high with no significant differences between groups. Neither fibronectin nor human plasma coating improved the adhesion of BMC to β-TCP in comparison to uncoated β-TCP. Since our clinical product consists of BMC in autologous serum, we also analyzed the adhesion of BMC that was cultured in DMEM medium without serum to evaluate the effect of the serum component on BMC. Adhesion of BMC cultured in serum-free medium and serum-containing medium did not differ significantly (Figures 1(a) and 1(b)). Pertaining to the selective enrichment or depletion of progenitor cells in the supernatant, the progenitor cell concentration in the initial BMC preparation and in the fraction of BMC that did not adhere to the biomaterials was evaluated by flow cytometry. No significant differences were observed (Figure 1(c)). This result indicates that neither surface coating nor medium conditions induced a selective enrichment or depletion of progenitor cell species on the β-TCP granules (Figure 1(c)).


Characterization of bone marrow mononuclear cells on biomaterials for bone tissue engineering in vitro.

Henrich D, Verboket R, Schaible A, Kontradowitz K, Oppermann E, Brune JC, Nau C, Meier S, Bonig H, Marzi I, Seebach C - Biomed Res Int (2015)

Seeding efficacy of BMC depending on the surface coating and medium conditions. BMCs (n = 16) were seeded on β-TCP either coated with fibronectin or human fresh frozen plasma and on uncoated β-TCP in the presence of MesenCult medium or DMEM. Seeding efficacy (%) is plotted on the ordinate. No significant differences in seeding efficacy were observed (a). Representative fluorescence microscopy images of DAPI stained BMC seeded under the different conditions are shown in (b). Similar amounts of adhering BMC were seen under each condition. No selective enrichment or depletion of specific progenitor cell subpopulation was observed on the surface of all biomaterials in various medium conditions as shown in (c). The percentages of progenitor cell types which resemble EPC (CD34+/CD45+; CD34+/CD133+/CD45+) and MSC (CD45−/CD271+) were analyzed in the fraction of nonadhering cells. The similar values indicate that no selective enrichment of progenitor cell subpopulations occurred due to the different culture and surface conditions.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4352750&req=5

fig1: Seeding efficacy of BMC depending on the surface coating and medium conditions. BMCs (n = 16) were seeded on β-TCP either coated with fibronectin or human fresh frozen plasma and on uncoated β-TCP in the presence of MesenCult medium or DMEM. Seeding efficacy (%) is plotted on the ordinate. No significant differences in seeding efficacy were observed (a). Representative fluorescence microscopy images of DAPI stained BMC seeded under the different conditions are shown in (b). Similar amounts of adhering BMC were seen under each condition. No selective enrichment or depletion of specific progenitor cell subpopulation was observed on the surface of all biomaterials in various medium conditions as shown in (c). The percentages of progenitor cell types which resemble EPC (CD34+/CD45+; CD34+/CD133+/CD45+) and MSC (CD45−/CD271+) were analyzed in the fraction of nonadhering cells. The similar values indicate that no selective enrichment of progenitor cell subpopulations occurred due to the different culture and surface conditions.
Mentions: The seeding efficacy was generally high with no significant differences between groups. Neither fibronectin nor human plasma coating improved the adhesion of BMC to β-TCP in comparison to uncoated β-TCP. Since our clinical product consists of BMC in autologous serum, we also analyzed the adhesion of BMC that was cultured in DMEM medium without serum to evaluate the effect of the serum component on BMC. Adhesion of BMC cultured in serum-free medium and serum-containing medium did not differ significantly (Figures 1(a) and 1(b)). Pertaining to the selective enrichment or depletion of progenitor cells in the supernatant, the progenitor cell concentration in the initial BMC preparation and in the fraction of BMC that did not adhere to the biomaterials was evaluated by flow cytometry. No significant differences were observed (Figure 1(c)). This result indicates that neither surface coating nor medium conditions induced a selective enrichment or depletion of progenitor cell species on the β-TCP granules (Figure 1(c)).

Bottom Line: On day 14, metabolic activity was significantly increased in BMC seeded on DBM in comparison to BMC seeded on BS.Likewise increased VEGF-synthesis was observed on day 2 in BMC seeded on DBM when compared to BMC seeded on BS.Beta-TCP and DBM were similar and both superior to BS, suggesting either as suitable materials for spatial restriction of BMC used for regenerative medicine purposes in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Trauma, Hand and Reconstructive Surgery, Hospital of the Goethe University, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany.

ABSTRACT
Bone marrow mononuclear cells (BMCs) are suitable for bone tissue engineering. Comparative data regarding the needs of BMC for the adhesion on biomaterials and biocompatibility to various biomaterials are lacking to a large extent. Therefore, we evaluated whether a surface coating would enhance BMC adhesion and analyze the biocompatibility of three different kinds of biomaterials. BMCs were purified from human bone marrow aspirate samples. Beta tricalcium phosphate (β-TCP, without coating or coated with fibronectin or human plasma), demineralized bone matrix (DBM), and bovine cancellous bone (BS) were assessed. Seeding efficacy on β-TCP was 95% regardless of the surface coating. BMC demonstrated a significantly increased initial adhesion on DBM and β-TCP compared to BS. On day 14, metabolic activity was significantly increased in BMC seeded on DBM in comparison to BMC seeded on BS. Likewise increased VEGF-synthesis was observed on day 2 in BMC seeded on DBM when compared to BMC seeded on BS. The seeding efficacy of BMC on uncoated biomaterials is generally high although there are differences between these biomaterials. Beta-TCP and DBM were similar and both superior to BS, suggesting either as suitable materials for spatial restriction of BMC used for regenerative medicine purposes in vivo.

No MeSH data available.


Related in: MedlinePlus