Microbubble-enhanced ultrasound for gene transfer into living skin equivalents

Yang, Lüjun; Shirakata, Yuji; Tamai, Katsuto; Dai, Xiuju; Hanakawa, Yasushi; Tokumaru, Sho; Yahata, Yoko; Tohyama, Mikiko; Shiraishi, Ken; Nagai, Hiroshi; Wang, Xiaoling; Murakami, Shinji; Sayama, Koji; Kaneda, Yasufumi; Hashimoto, Koji

doi:10.1016/j.jdermsci.2005.07.001

Cited by 36 publications

(25 citation statements)

References 42 publications

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“…The number of basal cells on dermal-epidermal junction is significantly higher than that of a conventional LSE made by seeding keratinocytes on fibroblast-populated type I collagen gel (Fig. 3B) [32,33]. The spinous layer, granular layer, and stratum corneum were also clearly identified.…”

Section: A Well Differentiated Epidermis Formed On De-epithelialized mentioning

confidence: 88%

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Living skin equivalents constructed using human amnions as a matrix

Yang¹,

Shirakata²,

Tokumaru

et al. 2009

Journal of Dermatological Science

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Section: A Well Differentiated Epidermis Formed On De-epithelialized mentioning

confidence: 88%

“…The keratinocytes were kept submerged in culture for 2 days. Then, the LSE was lifted to provide an air-liquid interface and cornification medium (1:1 mixture of Ham's F-12 and DMEM supplemented with 2% FCS and other supplements, as described previously [33]) was added. This medium was changed every other day.…”

Section: Preparation Of Skin Equivalentsmentioning

confidence: 99%

Living skin equivalents constructed using human amnions as a matrix

Yang¹,

Shirakata²,

Tokumaru

et al. 2009

Journal of Dermatological Science

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“…The preparation of collagen gel and LSE has been described previously [12]. Briefly, a collagen gel was prepared by mixing 6 volumes of ice-cold porcine collagen type I solution (Nitta Gelatin, Osaka, Japan) with 1 volume of 8 × DMEM (Gibco), 10 volumes of 1 × DMEM supplemented with 20% FCS, and 1 volume of 0.1 N NaOH.…”

Section: Methodsmentioning

confidence: 99%

“…The keratinocytes were kept submerged in culture medium for 2 days. When the keratinocytes reached confluence, the LSE was lifted to provide an air-liquid interface, and cornification medium composed of a 1: 1 mixture of Ham’s F-12 and DMEM supplemented with 2% FCS and other supplements [12] was added. This medium was changed every other day until 3 weeks after airlift.…”

Section: Methodsmentioning

confidence: 99%

Basic Fibroblast Growth Factor Influences Epidermal Homeostasis of Living Skin Equivalents through Affecting Fibroblast Phenotypes and Functions

Yang¹,

Zhang²,

Wu³

et al. 2018

Skin Pharmacol Physiol

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Aims: To elucidate the possible mechanisms of how basic fibroblast growth factor (bFGF) influences epidermal homeostasis in a living skin equivalent (LSE) model. Methods: Several wound healing-related growth factors were analyzed at protein and mRNA levels for dermal fibroblasts of induced alpha-smooth muscle actin (α-SMA)-positive or α-SMA-negative phenotypes. During culturing an LSE model by seeding normal human keratinocytes on a fibroblast-populated type I collagen gel, bFGF or neutralizing antibody for keratinocyte growth factor (KGF) was added to investigate its effects on fibroblast phenotypes and, subsequently, epidermal homeostasis by histology and immunohistochemistry. Results: The α-SMA-positive phenotype of fibroblasts induced by transforming growth factor beta-1 (TGF-β₁) markedly suppressed the expression of KGF and hepatocyte growth factor (HGF), and slightly upregulated vascular endothelial growth factor (VEGF) and TGF-β₁ at mRNA and protein levels, compared with α-SMA-negative fibroblasts treated with bFGF. α-SMA expression of fibroblasts at the epidermal-mesenchymal junction of the LSEs was suppressed by the addition of bFGF, and a better-differentiated epidermis was presented. The abrogation of KGF from fibroblasts by the addition of the KGF neutralizing antibody disenabled the LSE culturing system to develop an epidermis. Conclusions: bFGF, through affecting the phenotypes and functions of fibroblasts, especially KGF expression, influenced epidermal homeostasis in an LSE model.

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“…Only the long-term presence of the transfected gene in the tendons can continuously produce cytokines and promote healing. Yang et al 29,30 indicated that genes transfected using UTMD could exist in different tissues for weeks or even 6 months or more. In our study, although the GFP expression was gradually decrease over time, on 56 days after local tendon injection, fluorescence expression was still visible in the Achilles tendons.…”

Section: Length Of Egfp Expression In the Achilles Tendonmentioning

confidence: 99%

Ultrasound-targeted microbubble destruction enhances naked plasmid DNA transfection in rabbit Achilles tendons in vivo

et al. 2011

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The study was to investigate the probability of increasing the transfection of the gene in tendons by ultrasound-targeted microbubble destruction (UTMD), and to search for the most suitable transfection conditions. A mixture of microbubbles and enhanced green fluorescent protein (EGFP) plasmids was injected into rabbit Achilles tendons by different administration routes and the tendons were ultrasound pulse by different ultrasonic conditions in order to determine the most appropriate conditions. Then, the rabbits were divided into four groups: (1) ultrasound + microbubbles + plasmid; (2) ultrasound+ plasmid; (3) microbubble + plasmid; (4) plasmid only. EGFP expression in the tendons and other tissues, and the damage to tendon and paratenon were all observed. The results showed that EGFP expression in the tendon was higher by ultrasound pulse with 2 W cm À2 of output intensity and a 20% duty cycle for 10 min. Local injection was determined to be the better administration route. Among the four groups, EGFP expression in Group 1 was higher than that in other groups. EGFP expression was highest on seventh day, then it gradually decrease over time, and lasted more than 56 days. EGFP expression was not found in other tissues. There was no obvious injury caused by UTMD. Under suitable conditions, it is feasible to use UTMD as a safe and effective gene transfection therapy for tendon injuries.

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Microbubble-enhanced ultrasound for gene transfer into living skin equivalents

Cited by 36 publications

References 42 publications

Living skin equivalents constructed using human amnions as a matrix

Living skin equivalents constructed using human amnions as a matrix

Basic Fibroblast Growth Factor Influences Epidermal Homeostasis of Living Skin Equivalents through Affecting Fibroblast Phenotypes and Functions

Ultrasound-targeted microbubble destruction enhances naked plasmid DNA transfection in rabbit Achilles tendons in vivo

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