Bioalternatives

Thanks to its expertise in cellular and tissue engineering, Bioalternatives has developed in vitro or ex vivo study models that are suitable for analyzing the interactions between microbiota and skin:

• Isolated bacterial culture (Staphylococcus aureus, Staphylococcus epidermidis, Propionibacterium acnes)
  
• infection experiments with the HSV-1 virus ( Herpes simplex virus type I)
  

Co-culture:
- bacteria on 2D skin models (keratinocytes, fibroblasts, sebocytes, etc.)
- bacteria on 3D skin models (explants, reconstructed epidermis, etc.)

Both the impact of compounds or formulations on the development of bacteria and the influence of bacteria on cutaneous response can be assessed by these models.

Bioalternatives offers a panel of solutions for the evaluation of:

• Bacterial viability
  
• Quantification of bacterial colony (CFU)
  
• Determination of bacterial load (qPCR (Taqman) or viral load
  
• Bacterial adhesion on 3D models (explants, reconstucted epidermis)
  
• Antimicrobial activity of compounds or formulations
  
• Biological response of 2D or 3D models (differentiation, inflammation and immune response)

Drawing from years of experience in cell and tissue engineering, BIOalternatives uses validated healthy cell and tissue systems to develop relevant models that mimic normal physiology, pathologies, inflammation, ageing, etc. These in vitro models are suitable for:
- Biological target identification
- Hit to lead selection
- Drug candidate profiling
- Mechanism of action & proof-of-concept studies

BIOalternatives immortalized a human sebocyte cell line (SEBO662), which responds to lipogenic stimulations and which can be used to select potential inhibitors. This cell line was genetically modified in order to express a functional receptor to androgens (AR), as in the sebaceous gland. In this model, active androgens (dihydrotestosterone (DHT) or testosterone), tested alone in a non-supplemented basal medium, induce strong specific responses including the expression of androgen-sensitive genes but also the induction of a sebocyte differentiation program and lipid synthesis and storage.
From this work resulted a wide range of new cell based assays useful for the selection and evaluation of anti-androgen compounds or pro and anti-seborrheic compounds (acne, oily skin, dry skin, alopecia and hirsutism).

in vitro pharmacological solutions for the testing of atopic dermatitis therapies

Atopic dermatitis is an inflammatory skin disease that is pruritic and recurrent. BIOalternatives research on atopic dermatitis has been aiming at the development of in vitro models such as inflamed keratinocytes and activated immune cells. We offer various standard assays to select atopic dermatitis targeting compounds and to characterize their effects at the major stages of the physiopathology: initiation, immune response, skin lesions and antimicrobial defenses.

in vitro pharmacological solutions for the testing of psoriasis therapies

BIOalternatives research on psoriasis has been aiming at the development of in vitro models of keratinocyte inflammation. After analyzing the effects of conditioned media from LT clones on keratinocytes, we have analyzed the cytokine content of these media and screened a library of more than 40 cytokines for their responsiveness in keratinocyte cultures and also screened cytokine associations. These studies led to the identification of a cytokine mix (association of 5 cytokines; Patent # WO2006063864 A3) that induces a strong synergistic effect on keratinocytes in culture.
BIOalternatives offers various standard assays to select psoriasis targeting compounds and to characterize their effects at the major stages of the physiopathology.

in vitro pharmacology solutions for the testing of wound care therapies

Wound healing is a complex and dynamic process of restoring skin cellular structures and tissue layers. BIOalternatives offers a range of in vitro models and assays to select active ingredients and to characterize their effects at the major stages of the physiopathology: hemostasis, inflammatory response, cell proliferation and granulation tissue formation and skin remodeling.

Are you looking for solutions to support your claims? in vitro assays are highly effective for establishing proof-of-concept of your compounds (e.g. extracts, active ingredients, formulations).

BIOalternatives, a premier in vitro dermatological testing service laboratory, can assist you at every step of the pre-clinical development of your products: screening of bioactive compounds, proof-of-concept studies, repositioning of active ingredients.

Our knowledge of skin biology and its application to new target assessment, mechanistic
understanding and claim substantiation are without question of the highest standards in the industry :
- Epidermal regeneration
- Skin barrier and hydration
- Skin pigmentation
- Skin aging
- Skin protection
- Skin vascularization
- Skin inflammation
- Sebaceous gland regulation
- Adipocyte metabolism
- Hair regeneration

We provide a wide range validated assays using 3D-skin related models: dermis equivalent, reconstructed human epidermis, full thickness skin model, etc. for compound (ingredient or finish products) testing.

BIOalternatives developed and validated in vitro cell and tissue models (e.g. normal or aged dermal fibroblasts, full thickness skin) and assays for investigating the anti-aging effect of cosmetics compounds, such as:
- cell migration, proliferation or differentiation assays
- extracellular matrix synthesis or degradation assays (e.g. collagen or tropoelastin synthesis/ expression, MMP activities)
- gene expression assays.

BIOalternatives developed and validated an ex-vivo model of human hair follicle suitable for investigating the effect of compounds on:
• the stimulation/ inhibition of hair growth,
• the survival rate increase in cell cultures (anti apoptosis, anti-hair loss focus),
• The hair pigmentation.

These parameters can be analyzed by morphometric techniques (hair elongation and viability measurement) but also through complex molecular approaches: gene expression (dedicated PCRarrays or full genome profile, immunohistochemistry) or histology as special staining e.g. Fontana Masson for pigmentation.

Cell and tissue engineering, development of new comple in vitro mammalian models:
- Tissue microdissection
- Primary cell isolation
- Cell line construction
- Co-culture model system
- Organotypic explant culture
- Three dimensional reconstruction
- Phenotypic and functional characterization

Cellular and molecular pharmacology assays for hit-to-lead selection of active pharmaceutical ingredients and claim substantiation of healthcare products:
- cell-based assays
- Screening
- Profiling
- Proof-of-concept
- Mechanism of action
- Benchmarking
- Safety
- Testing of formulations prior clinical trial

Services for the development, customization and optimization of innovative research methods and protocols :
- Literature review
- Feasability study
- Protocol design
- Method validation
- Multiplexing, miniaturization and optimization

Biological sample collection, processing, storage and information management:
- Study design
- Sample Collection
- Method develoment and validation (tissue microarrays, cDNArrays, PCRarrays, immunoassays, biochemical assays)
- Analysis service
- Biobanking (RNA, cDNA, cells, frozen tissues, embedded tissues)
- Data management

BrdU Cell Proliferation assay or 3H thymidine incorporation assay

Cell migration is commonly assessed using the scratch assay which measures the movement of cells into an artificial wound made on a confluent cell monolayer to create a cell-free area. As an alternative to the scratch assay which has high variability, the Oris™ Cell Migration Assay allows the formation of homogeneously sized cell-free areas into which migration can occur without releasing factors from dead cells or damaging the underlying extracellular matrix.

Cell-Based Gene Expression Assays using qPCR (SYBR Green or TaqMan) or microrrays (Affymetrix);