Infused demineralized bone fibers

A method of making infused at least partially demineralized or at least partially decalcified bone fibers, comprising:

• (a) cutting or shaving whole bone into bone fibers;
• (b) washing the bone fibers;
• (c) at least partially demineralizing or at least partially decalcifying the bone fibers, creating a surface demineralization or decalcification that extends, into the bone fibers;
• and
• (d) infusing the at least partially demineralized or at least partially decalcified bone fibers with a supernatant of biologic material, a polyampholyte cryoprotectant, and a buffy coat layer comprising whole cells to create infused at least partially demineralized or at least partially decalcified bone fibers, wherein infusing comprises one or more of:
• i. exposing the at least partially demineralized or at least partially decalcified bone fibers, supernatant, polyampholyte cryoprotectant, and buffy coat layer to a negative pressure or vacuum to draw the supernatant, polyampholyte cryoprotectant, and buffy coat layer into the bone fibers;
• and
• ii. exposing the at least partially demineralized or at least partially decalcified bone fibers, supernatant, polyampholyte cryoprotectant, and buffy coat layer to a positive pressure to drive the supernatant, polyampholyte cryoprotectant, and buffy coat layer into the bone fibers.

A method of making an infused bone fiber mixture comprises:

• (a) at least partially demineralizing or at least partially decalcifying whole bone;
• (b) cutting or shaving the at least partially demineralized or at least partially decalcified whole bone into bone fibers, wherein the cut or shaved bone fibers form a bone fiber mixture comprising one or more of fully decalcified or fully demineralized bone fibers, at least partially decalcified or at least partially demineralized bone fibers, or mineralized and calcified bone fibers;
• (c) washing the bone fiber mixture;
• and
• (d) infusing the bone fiber mixture with a supernatant of biologic material, polyampholyte cryoprotectant, and buffy coat layer comprising whole cells to create an infused bone fiber mixture, wherein infusing comprises:
• i. exposing the bone fiber mixture, supernatant, polyampholyte cryoprotectant, and buffy coat layer to a negative pressure or vacuum to draw the supernatant, polyampholyte cryoprotectant, and buffy coat layer into the bone fibers;
• or
• ii. exposing the bone fiber mixture, supernatant, polyampholyte cryoprotectant, and buffy coat layer to a positive pressure to drive the supernatant, polyampholyte cryoprotectant, and buffy coat layer into the bone fibers.