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Millicurrent stimulation of human articular chondrocytes cultivated in a collagen type-I gel and of human osteochondral explants.

Gavénis K, Andereya S, Schmidt-Rohlfing B, Mueller-Rath R, Silny J, Schneider U - BMC Complement Altern Med (2010)

Bottom Line: Following millicurrent treatment, IL-6, TNFalpha, and MMP13 gene expression decreased.In general, the standard deviations of the gene expression data were high, resulting in rarely significant results.We conclude that millicurrent stimulation of human osteoarthritic chondrocytes cultivated in a 3 D collagen gel and of osteochondral explants directly influences cell metabolism.

View Article: PubMed Central - HTML - PubMed

Affiliation: Aachen University Hospital, Dept of Orthopaedic Surgery, Pauwelsstr 30, 52074 Aachen, Germany. kgavenis@ukaachen.de

ABSTRACT

Background: Here we investigate the effect of millicurrent treatment on human chondrocytes cultivated in a collagen gel matrix and on human osteochondral explants.

Methods: Human chondrocytes from osteoarthritic knee joints were enzymatically released and transferred into a collagen type-I gel. Osteochondral explants and cell-seeded gel samples were cultivated in-vitro for three weeks. Samples of the verum groups were stimulated every two days by millicurrent treatment (3 mA, sinusoidal signal of 312 Hz amplitude modulated by two super-imposed signals of 0.28 Hz), while control samples remained unaffected. After recovery, collagen type-I, type-II, aggrecan, interleukin-1beta, IL-6, TNFalpha and MMP13 were examined by immunohistochemistry and by real time PCR.

Results: With regard to the immunostainings 3 D gel samples and osteochondral explants did not show any differences between treatment and control group. The expression of all investigated genes of the 3 D gel samples was elevated following millicurrent treatment. While osteochondral explant gene expression of col-I, col-II and Il-1beta was nearly unaffected, aggrecan gene expression was elevated. Following millicurrent treatment, IL-6, TNFalpha, and MMP13 gene expression decreased. In general, the standard deviations of the gene expression data were high, resulting in rarely significant results.

Conclusions: We conclude that millicurrent stimulation of human osteoarthritic chondrocytes cultivated in a 3 D collagen gel and of osteochondral explants directly influences cell metabolism.

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Real-time PCR analysis of osteochondral explants. (a) Gene expression of various genes of matrix synthesis, turnover and inflammation of osteochondral explants left untreated (left column) or stimulated by millicurrent treatment (right column). Specimens were cultivated in-vitro for 3 weeks. The housekeeping-gene β-actin was used as internal standard. P = 0.05. (b) Calculated relative gene expression of treatment samples in relation to control samples of the same patients, where gene expression of control samples was set as 1.
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Figure 5: Real-time PCR analysis of osteochondral explants. (a) Gene expression of various genes of matrix synthesis, turnover and inflammation of osteochondral explants left untreated (left column) or stimulated by millicurrent treatment (right column). Specimens were cultivated in-vitro for 3 weeks. The housekeeping-gene β-actin was used as internal standard. P = 0.05. (b) Calculated relative gene expression of treatment samples in relation to control samples of the same patients, where gene expression of control samples was set as 1.

Mentions: In contrast to semi-quantitative detection of protein production, determination of gene expression revealed some differences. The absolute and relative gene expression data are given in Fig. 5. While gene expression of collagen type-I and type-II was nearly unaffected, aggrecan gene expression was elevated by 1.75 fold (p = 0.45) in treatment samples with respect to control samples of the same patients. IL1β gene expression showed an equivalent level in both control and treatment group, while IL-6 gene expression in the treatment group decreased to 0.22 (p = 0.16). Treatment group MMP-13 gene expression decreased to 0.55 (p = 0.19), and TNFα decreased to 0.48 (p = 0.87) relative to control samples of the same patients.


Millicurrent stimulation of human articular chondrocytes cultivated in a collagen type-I gel and of human osteochondral explants.

Gavénis K, Andereya S, Schmidt-Rohlfing B, Mueller-Rath R, Silny J, Schneider U - BMC Complement Altern Med (2010)

Real-time PCR analysis of osteochondral explants. (a) Gene expression of various genes of matrix synthesis, turnover and inflammation of osteochondral explants left untreated (left column) or stimulated by millicurrent treatment (right column). Specimens were cultivated in-vitro for 3 weeks. The housekeeping-gene β-actin was used as internal standard. P = 0.05. (b) Calculated relative gene expression of treatment samples in relation to control samples of the same patients, where gene expression of control samples was set as 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2921357&req=5

Figure 5: Real-time PCR analysis of osteochondral explants. (a) Gene expression of various genes of matrix synthesis, turnover and inflammation of osteochondral explants left untreated (left column) or stimulated by millicurrent treatment (right column). Specimens were cultivated in-vitro for 3 weeks. The housekeeping-gene β-actin was used as internal standard. P = 0.05. (b) Calculated relative gene expression of treatment samples in relation to control samples of the same patients, where gene expression of control samples was set as 1.
Mentions: In contrast to semi-quantitative detection of protein production, determination of gene expression revealed some differences. The absolute and relative gene expression data are given in Fig. 5. While gene expression of collagen type-I and type-II was nearly unaffected, aggrecan gene expression was elevated by 1.75 fold (p = 0.45) in treatment samples with respect to control samples of the same patients. IL1β gene expression showed an equivalent level in both control and treatment group, while IL-6 gene expression in the treatment group decreased to 0.22 (p = 0.16). Treatment group MMP-13 gene expression decreased to 0.55 (p = 0.19), and TNFα decreased to 0.48 (p = 0.87) relative to control samples of the same patients.

Bottom Line: Following millicurrent treatment, IL-6, TNFalpha, and MMP13 gene expression decreased.In general, the standard deviations of the gene expression data were high, resulting in rarely significant results.We conclude that millicurrent stimulation of human osteoarthritic chondrocytes cultivated in a 3 D collagen gel and of osteochondral explants directly influences cell metabolism.

View Article: PubMed Central - HTML - PubMed

Affiliation: Aachen University Hospital, Dept of Orthopaedic Surgery, Pauwelsstr 30, 52074 Aachen, Germany. kgavenis@ukaachen.de

ABSTRACT

Background: Here we investigate the effect of millicurrent treatment on human chondrocytes cultivated in a collagen gel matrix and on human osteochondral explants.

Methods: Human chondrocytes from osteoarthritic knee joints were enzymatically released and transferred into a collagen type-I gel. Osteochondral explants and cell-seeded gel samples were cultivated in-vitro for three weeks. Samples of the verum groups were stimulated every two days by millicurrent treatment (3 mA, sinusoidal signal of 312 Hz amplitude modulated by two super-imposed signals of 0.28 Hz), while control samples remained unaffected. After recovery, collagen type-I, type-II, aggrecan, interleukin-1beta, IL-6, TNFalpha and MMP13 were examined by immunohistochemistry and by real time PCR.

Results: With regard to the immunostainings 3 D gel samples and osteochondral explants did not show any differences between treatment and control group. The expression of all investigated genes of the 3 D gel samples was elevated following millicurrent treatment. While osteochondral explant gene expression of col-I, col-II and Il-1beta was nearly unaffected, aggrecan gene expression was elevated. Following millicurrent treatment, IL-6, TNFalpha, and MMP13 gene expression decreased. In general, the standard deviations of the gene expression data were high, resulting in rarely significant results.

Conclusions: We conclude that millicurrent stimulation of human osteoarthritic chondrocytes cultivated in a 3 D collagen gel and of osteochondral explants directly influences cell metabolism.

Show MeSH