Preface

We recommend Bio incubation as an advanced method for preparing PRF that uses controlled heat to create a flowable, injectable fibrin matrix with enhanced regenerative and anti-inflammatory properties. Each PRF preparation method offers unique clinical advantages and limitations, and the choice depends on the specific therapeutic application and available equipment.

Bio incubation is a preferred method for PRF preparation in aesthetics because it produces a safe, natural, and injectable gel that combines the benefits of immediate volumization with long-term tissue regeneration. This method maximizes patient safety, satisfaction, and clinical outcomes in facial rejuvenation and contouring 1,2,7.

Several protocols and technologies exist for preparing PRF, each affecting the composition, physical properties, and clinical applications of the final product. Below are the main methods, including the emerging technique of bio incubation, along with their advantages and disadvantages.

Bio Incubation PRF (Heated/Bio-Filler PRF)

Method:

• After PRP or liquid PRF is prepared, it is incubated in a controlled heating device (bio incubator), then cooled 2,5.
• Heat triggers the release of calcium from platelets, activating the thrombotic cascade and forming a dense, flowable fibrin matrix 2,5.
• The product is then cooled to stabilize the matrix and maintain flowability 2.

Advantages:

• Produces a flowable, gel-like PRF suitable for injection and volume restoration (bio-filler) 2,5.
• Heat-induced activation is gentler than chemical activation (e.g., CaCl₂ or bovine thrombin), reducing pro-inflammatory byproducts and patient discomfort 2.
• Enhanced anti-inflammatory properties and stable for hours after preparation 2,5.
• Complies with FDA minimal manipulation guidelines 2.

Disadvantages:

• Requires specialized bio incubator and cooling equipment 2,5.
• Additional steps add complexity and cost.
• Still relatively new; long-term comparative data is limited.

Classic PRF (Leukocyte- and Platelet-Rich Fibrin, L-PRF)

Method:

Blood is drawn into glass tubes without anticoagulant and centrifuged rapidly (e.g., 750g for 12 min) 4.

Platelets and leukocytes become entrapped in the fibrin clot as it forms naturally.

Advantages:

• Simple, cost-effective, no chemical additives required 1,4.
• Produces a solid, dense fibrin matrix rich in platelets and leukocytes, supporting robust tissue healing 1.
• Growth factors are released slowly over up to 14 days 4.

Disadvantages:

• Must process blood quickly (within 2 minutes) to avoid premature clotting 1,4.
• Produces a solid clot, not suitable for injection into fine tissues.

Injectable PRF (I-PRF)

Method:

• Very low centrifugation speed (e.g., 700 rpm for 3 min), yielding a liquid PRF 8.
• Can be injected before polymerization occurs.

Advantages:

• Injectable, allowing use in soft tissue, joints, facial aesthetics, and hair restoration 8.
• High concentrations of platelets, leukocytes, and growth factors.

Disadvantages:

• Must be used immediately before it gels.
• Short working time 7-8 minutes, technique-sensitive.

Horizontal PRF (H-PRF, Bio-PRF)

Method:

• Uses a horizontal centrifuge to ensure even cell distribution (e.g., Bio-PRF system) 3,6.
• Can accumulate up to 4 times more platelets and leukocytes than fixed-angle centrifugation 3,6.

Advantages:

• More homogeneous and higher concentration of platelets and leukocytes 3,6.
• Potentially superior regenerative outcomes.

Disadvantages:

• Requires specialized horizontal centrifuge equipment.
• Less widely available.

More on bio incubation

100% Natural, Additive-Free Filler

Bio incubation uses only the patient’s own blood, processed without any chemical additives or anticoagulants. This eliminates the risk of allergic reactions or synthetic material complications, making it one of the safest options for skin rejuvenation and volume restoration 1,7.

Creation of a Flowable, Injectable Gel

Traditional PRF rapidly clots, limiting its use as a dermal filler. Bio incubation involves gently heating the plasma (typically to 42–47°C for several minutes), which induces polymerization of fibrinogen into a gel-like matrix. This process produces a flowable, growth factor-rich gel—sometimes called PRF BioFiller or ezGEL—that can be injected much like a conventional filler to restore facial volume, contour, and smooth lines 7.

Sustained Growth Factor Release

The heat-induced fibrin network formed during bio incubation traps platelets and growth factors, which are then released gradually over time. This sustained release is critical for stimulating ongoing collagen and elastin production, leading to long-term improvements in skin texture, firmness, and elasticity—key goals in facial aesthetics 2,7.

Enhanced Collagen and Elastin Stimulation

PRF BioFiller acts as both a volumizer and a biostimulator. The injected gel not only restores lost facial volume but also stimulates the body’s own collagen and elastin synthesis, leading to progressive, natural rejuvenation beyond the initial treatment window 1,7.

Minimal Downtime and Side Effects

Because the process is autologous and additive-free, patients experience minimal swelling, bruising, or downtime. Most can resume normal activities quickly, making it highly attractive for busy individuals seeking subtle, natural-looking enhancements 1,7.

Versatility and Personalization

Bio incubation allows clinicians to tailor the viscosity and handling properties of the PRF gel, making it suitable for a variety of facial areas—under eyes, cheeks, nasolabial folds, lips, temples, and more. The treatment is personalized to each patient’s needs and biology 1,7.

Long-Lasting, Natural Results

Unlike traditional fillers that may dissipate or migrate, PRF BioFiller integrates with native tissue and supports ongoing tissue regeneration. Results are both immediate (due to the gel’s volume) and progressive (as collagen remodeling occurs), with improvements that can last for months 7.

References

1 Platelet-rich fibrin: Basics of biological actions and protocol (PMC7985567)

2 Optimizing Heat Process for Superior Platelet-Rich Fibrin (YouTube, 2025)

3 Bio-PRF: Home (bio-prf.com)

4 Platelet-Rich Fibrin, Preparation and Use in Dermatology (PMC8664174)

5 Platelet Rich Fibrin Bio-Filler - Juventix Regenerative Medical

6 Science Behind PRF | Bio-PRF - Emerginnova

7 Impact of incubation method on the release of growth factors in non-activated PRF (ScienceDirect)

8 Science Behind Bio-PRF (bio-prf.com)